Use of lipid conjugates in the treatment of diseases or disorders of the eye

ABSTRACT

In one embodiment, the invention provides a method of treating, reducing the incidence, reducing the severity or pathogenesis of an eye disease or disorder in a subject, including, inter alia, retinal detachment, macular degeneration, glaucoma or retinopathy, comprising the step of administering an effective amount of a lipid or phospholipid moiety bound optionally via a spacer to a physiologically acceptable monomer, dimer, oligomer, or polymer via an ester or amide bond, and/or a pharmaceutically acceptable salt or a pharmaceutical product thereof. This invention also provides a contact lens solution comprising a lipid or phospholipid moiety bound optionally via a spacer to a physiologically acceptable monomer, dimer, oligomer, or polymer via an ester or amide bond, and/or a pharmaceutically acceptable salt or a pharmaceutical product thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is continuation of U.S. application Ser. No.11/984,223, filed Nov. 14, 2007, which claims the benefit of U.S.Provisional Application Ser. No. 60/858,706, filed Nov. 14, 2006 andU.S. Provisional Application Ser. No. 60/907,785, filed Apr. 17, 2007.All applications above are incorporated herein in their entirety.

FIELD OF THE INVENTION

This invention provides compounds and methods of use thereof forreducing the incidence, reducing the severity or pathogenesis, ortreating a disease or disorder of the eye in a subject, including, interalia, retinal detachment, macular degeneration, glaucoma or retinopathy,as well as contact lens solutions comprising said compounds.

BACKGROUND OF THE INVENTION

Compounds for use in the present invention are thought to inhibit theenzyme phospholipase A2 (PLA2, EC 3.1.1.4). Phospholipase A2 catalyzesthe breakdown of phospholipids at the sn-2 position to produce a fattyacid and a lysophospholipid. The activity of this enzyme has beencorrelated with various cell functions, particularly with the productionof lipid mediators such as eicosanoid production (prostaglandins,thromboxanes and leukotrienes), platelet activating factor andlysophospholipids. Compounds for use in the present invention may offera wider scope of protection of cells and organisms from injurious agentsand pathogenic processes, including the prevention and treatment of eyediseases.

The elderly population in the United States is increasing rapidly. Bythe year 2030, approximately 70 million Americans will be over 65 yearsof age. Loss of vision among the elderly is a major health care problem:approximately one in three elderly persons has some form ofvision-reducing eye disease by the age of 65. Vision impairment isassociated with a decreased ability to perform activities of dailyliving and an increased risk for depression.

Although estimates vary, there are approximately 10 million blind andvisually impaired people in the United States, of which approximately5.5 million are elderly individuals. Cataract, glaucoma, age-relatedmacular degeneration, and diabetic retinopathy are the four eyedisorders that pose the greatest threats to vision after age 40.Patients with age-related macular degeneration often have the followingsymptoms: blurred vision, image distortion, central scotoma, and/ordifficulty reading; Patients with glaucoma often have the followingsymptoms: visual field loss and/or blurred vision (late); Patients withcataracts often complain of blurred vision, glare, and/or monoculardiplopia; Patients with diabetic retinopathy often have the followingsymptoms: Bluffed vision, floaters, visual field loss, often have poornight vision.

Other common eye disorders of aging, include presbyopia, dry eye,floaters and flashes, retinal detachment, and eyelid problems such asdrooping upper or lower lids.

In the United States, diabetes is responsible for 8% of legal blindness,making it the leading cause of new cases of blindness in adults 20-74years of age. Each year, between 12,000 to 24,000 people lose theirsight because of diabetes, making patients with diabetes 25 times morelikely to lose vision than those who are not diabetic, according to theAmerican Academy of Ophthalmology. In addition, diabetic retinopathyoften leads to additional eye disorders such as retinal detachment,glaucoma cataract, and corneal disease, contributing to the high rate ofblindness in diabetics.

Over 29 million people in the United States wear contact lenses, whichprovide a safe and effective way to correct vision when used with careand proper supervision. However, many contact lens wearers and potentialcontact lens wearers suffer from discomfort, dry eyes, and infection asa result of contact lens use. There is therefore a need to designcontact lenses to comprise compounds that allow the lens to be morebiocompatible, comfortable, tear-wettable, anti-bacterial and oxygenpermeable.

SUMMARY OF THE INVENTION

In one embodiment, the invention provides a method of reducing theincidence, reducing the severity or pathogenesis of a disease ordisorder of the eye in a subject comprising the step of contacting saidsubject with a compound comprising a lipid or phospholipid moiety boundoptionally via a spacer to a physiologically acceptable monomer, dimer,oligomer, or polymer via an ester or amide bond, and/or apharmaceutically acceptable salt or a pharmaceutical product thereof. Inanother embodiment, the invention provides a method of treating adisease or disorder of the eye in a subject comprising the step ofcontacting said subject with a compound comprising a lipid orphospholipid moiety bound optionally via a spacer to a physiologicallyacceptable monomer, dimer, oligomer, or polymer via an ester or amidebond, and/or a pharmaceutically acceptable salt or a pharmaceuticalproduct thereof. In another embodiment, the invention provides a contactlens solution comprising a lipid or phospholipid moiety bound optionallyvia a spacer to a physiologically acceptable monomer, dimer, oligomer,or polymer via an ester or amide bond, and/or a pharmaceuticallyacceptable salt or a pharmaceutical product thereof.

In one embodiment, the compound for use in the solutions, compositionsand methods of the present invention is represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline,        phosphate, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000;    -   wherein any bond between L, Z, Y and X is either an amide or an        esteric bond.

In one embodiment, the compound for use in the solutions, compositionsand methods of the present invention is represented by the structure ofthe general formula (A):

wherein

-   L is a lipid or a phospholipid;-   Z is either nothing, ethanolamine, serine, inositol, choline,    phosphate, or glycerol;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is a physiologically acceptable monomer, dimer, oligomer, or    polymer; and-   n is a number from 2 to 1000;-   wherein any bond between L, Z, Y and X is either an amide or an    esteric bond.

In another embodiment, the compound for use in the solutions,compositions and methods of the present invention is represented by thestructure of the general formula (I):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is alginate, hydroxyethylstarch, polygeline,    carboxymethylcellulose, or a combination thereof; and-   n is a number from 1 to 1000-   or-   wherein-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is alginate, hydroxyethylstarch, polygeline,    carboxymethylcellulose, or a combination thereof; and-   n is a number from 1 to 1000.

In another embodiment, the compound for use in the solutions,compositions and methods of the present invention is represented by thestructure of the general formula (I):

wherein

-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is alginate, hydroxyethylstarch, polygeline,    carboxymethylcellulose, or a combination thereof; and-   n is a number from 2 to 1000-   or-   wherein-   R₁ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,    alkyl chain ranging in length from 2 to 30 carbon atoms;-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms;-   X is alginate, hydroxyethylstarch, polygeline,    carboxymethylcellulose, or a combination thereof; and-   n is a number from 2 to 1000.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment, the invention provides a method of reducing theincidence, reducing the severity or pathogenesis of a disease ordisorder of the eye in a subject comprising the step of contacting saidsubject with a compound comprising a lipid or phospholipid moiety boundoptionally via a spacer to a physiologically acceptable monomer, dimer,oligomer, or polymer via an ester or amide bond, and/or apharmaceutically acceptable salt or a pharmaceutical product thereof.

In one embodiment, the invention provides a method of suppressing orinhibiting, a disease or disorder of the eye in a subject, comprisingthe step of contacting a cell with a compound comprising a lipid orphospholipid moiety bound to a physiologically acceptable monomer,dimer, oligomer, or polymer, and/or a pharmaceutically acceptable saltor a pharmaceutical product thereof.

In another embodiment, the invention provides a method of treating adisease or disorder of the eye in a subject, comprising the step ofcontacting said subject with a compound comprising a lipid orphospholipid moiety bound optionally via a spacer to a physiologicallyacceptable monomer, dimer, oligomer, or polymer via an ester or amidebond, and/or a pharmaceutically acceptable salt or a pharmaceuticalproduct thereof.

In another embodiment, the invention provides a method of preventing adisease or disorder of the eye in a subject, comprising the step ofcontacting said subject with a compound comprising a lipid orphospholipid moiety bound optionally via a spacer to a physiologicallyacceptable monomer, dimer, oligomer, or polymer via an ester or amidebond, and/or a pharmaceutically acceptable salt or a pharmaceuticalproduct thereof.

In one embodiment, the invention provides for the use of a lipid orphospholipid moiety bound optionally via a spacer to a physiologicallyacceptable monomer, dimer, oligomer, or polymer via an ester or amidebond, in the preparation of a composition for suppressing, inhibiting,preventing or treating a disease or disorder of the eye in a subject. Inanother embodiment, the invention provides for the use of a lipid orphospholipid moiety bound optionally via a spacer to a physiologicallyacceptable monomer, dimer, oligomer, or polymer via an ester or amidebond, in the preparation of a composition for reducing the incidence,reducing the severity or pathogenesis of a disease or disorder of theeye in a subject.

In one embodiment, the term “a disease or disorder of the eye” refers toany one or more of the following conditions: retinal detachment, macularedema, retinopathy, age-related macular degeneration, macular cyst,macular hole, solar retinopathy, diabetic retinopathy, branch retinalvein occlusion, or Lebers congenital amaurosis. In another embodiment,the term “a disease or disorder of the eye” refers to any one or more ofthe following conditions: corneal graft rejection, uveitis, inflammatoryeye diseases, infectious eye diseases, ocular tumours, neovascularproliferative diseases, neovascular maculopathies, rheumatoid cornealmelting disorders, or autoimmune disorders.

It is to be understood that the method of the present invention may beused to prevent or treat any disorder or disease of the eye orassociated with the eye, or in another embodiment, any ophthalmicdisorder. In one embodiment, the methods of the present invention may beused to prevent, suppress, inhibit or treat episcleritis, scleritis, ora combination thereof. In another embodiment, the methods of the presentinvention may be used to prevent, suppress, inhibit or treatretinopathy, including, inter alia, diabetic retinopathy, glaucoma,macular degeneration, retinal detachment, or a combination thereof. Inanother embodiment, the methods of the present invention may be used toprevent, suppress, inhibit or treat any one or more of the followingdiseases or disorders, or symptoms as a result thereof:achromatopsia/Maskun, amblyopia, anisometropia, Argyll Robertson pupil,astigmatism, anisometropia, blindness, chalazion, color blindness,achromatopsia/Maskun, esotropia, exotropia, floaters, vitreousdetachment, Fuchs' dystrophy, hypermetropia, hyperopia, hypertensiveretinopathy, iritis, keratoconus, Leber's congenital amaurosis, Leber'shereditary optic neuropathy, macular edema, myopia, nyctalopia,ophthalmoplegia, including progressive external ophthalmoplegia andinternal opthalmoplegia, opthalmoparesis, presbyopia, pterygium, red eye(medicine), retinitis pigmentosa, retinopathy of prematurity,retinoschisis, river blindness, ophthalmoplegia, scotoma, snowblindness/arc eye, eyelid disorders, ptosis, extraocular tumours,strabismus, which in one embodiment is esotropias, exotropias, verticalpatterns, eye injuries, or a combination thereof. In another embodiment,the methods of the present invention may be used to prevent, suppress,inhibit or treat any one or more of the following diseases or disorders,or symptoms as a result thereof: neovascular glaucoma, retrolentalfibroplasias, Vitamin A deficiency, contact lens overwear, atopickeratitis, superior limbic keratitis, pterygium keratitis sicca,Sjogrens syndrome, acne rosacea, phylectenulosis, syphilis, lipiddegeneration, chemical burns, Mooren ulcer, Terrien's marginaldegeneration, marginal keratolysis, polyarteritis, trauma, Wegenerssarcoidosis, scleritis, Steven's Johnson disease, periphigoid radialkeratotomy, sickle cell anemia, syphilis, pseudoxanthoma elasticum,Paget's disease, vein occlusion, artery occlusion, carotid obstructivedisease, chronic uveitis/vitritis, Lyme disease, systemic lupuserythematosis, Eales disease, Behcet's disease, presumed ocularhistoplasmosis, Best's disease, optic pits, Stargardt's disease, parsplanitis, chronic retinal detachment, retinoschisis, hyperviscositysyndromes, toxoplasmosis, trauma, post-laser complications, rubeosis, ora combination thereof.

In another embodiment, the methods of the present invention may be usedin combination with or to prevent or treat secondary effects of:intraocular lens replacement; ophthalmic enucleation, evisceration,exenteration, or a combination thereof; lacrimal sac surgeries; cornealpterygium; lamellar keratoplasty; penetrating keratoplasty, or acombination thereof, as well as any of the disorders or conditionsmentioned herein.

In one embodiment, the disease or disorder of the eye affects theanterior region of the eye, while in another embodiment, it affects theposterior region of the eye, while in another embodiment, it affectsboth the anterior and posterior regions of the eye. In one embodiment,the anterior segment includes the cornea, anterior chamber, iris andciliary body (anterior choroid), posterior chamber and crystalline lensand the posterior segment includes the retina with optic nerve, choroid(posterior choroid) and vitreous. In one embodiment, eye disordersresulting from the pathologic conditions of structures in the anteriorsegment of the eye are dry eye syndrome, keratitis or corneal dystrophy,cataracts, and glaucoma. In one embodiment, the disease or disorders ofthe posterior segment of the eye in general are retinal or choroidalvascular diseases or hereditary diseases such as Lebers congenitalamaurosis.

In one embodiment, “treating” refers to both therapeutic treatment andprophylactic or preventive measures, wherein the object is to prevent orlessen the targeted pathologic condition or disorder as describedhereinabove. Thus, in one embodiment, treating may include suppressing,inhibiting, preventing, treating, or a combination thereof. Thus, in oneembodiment, “treating” refers, inter alia, to increasing time tosustained progression, expediting remission, inducing remission,augmenting remission, speeding recovery, increasing efficacy of ordecreasing resistance to alternative therapeutics, or a combinationthereof. In one embodiment, “preventing” refers, inter alia, to delayingthe onset of symptoms, preventing relapse to a disease, decreasing thenumber or frequency of relapse episodes, increasing latency betweensymptomatic episodes, or a combination thereof. In one embodiment,“suppressing” or “inhibiting”, refers, inter alia, to reducing theseverity of symptoms, reducing the severity of an acute episode,reducing the number of symptoms, reducing the incidence ofdisease-related symptoms, reducing the latency of symptoms, amelioratingsymptoms, reducing secondary symptoms, reducing secondary infections,prolonging patient survival, or a combination thereof.

In one embodiment, symptoms are primary, while in another embodiment,symptoms are secondary. In one embodiment, “primary” refers to a symptomthat is a direct result of an eye disease, while in one embodiment,“secondary” refers to a symptom that is derived from or consequent to aprimary cause. In one embodiment, the compounds for use in the presentinvention treat primary or secondary symptoms or secondary complicationsrelated to an eye disease. In another embodiment, the compounds for usein the present invention treat primary or secondary symptoms orsecondary complications related to an eye disease or disorder.

In another embodiment, “symptoms” may be any manifestation of a diseaseor pathological condition, comprising inflammation, swelling, fever,pain, bleeding, itching, runny nose, coughing, headache, migraine,dizziness, blurry vision, decreased visual acuity, light sensitivity,etc., or a combination thereof. In one embodiment, symptoms compriseitchy eyes, swollen eyelids, redness, irritation, watery eyes, mucoiddischarge, pain, or a combination thereof.

Thus, in one embodiment of the present invention, the compounds for usein the present invention are directed towards the resolution of symptomsof a disease or disorder of the eye. In another embodiment, thecompounds affect the pathogenesis underlying a disease or disorder ofthe eye.

In one embodiment, a disease or disorder of the eye may affect a cell,in one embodiment, a vertebrate cell, in another embodiment, a mammaliancell, and in another embodiment, a human cell. It is to be understoodthat compounds of the present invention may be efficacious in treatingany cell type in which a disease or disorder of the eye or the causes ofa disease or disorder of the eye may exert an effect. In one embodiment,a compound for use in the present invention may localize to or act on aspecific cell type. In one embodiment, a compound for use in the presentinvention may be cytoprotective. In one embodiment a compound for use inthe present invention may be inserted or partially inserted into a cellmembrane. In another embodiment a compound for use in the presentinvention may be effective in treating a plurality of cell types.

In one embodiment, a disease or disorder of the eye is a primary orsecondary symptom of an underlying illness, which in one embodiment, isan autoimmune disease. In one embodiment, the underlying illness isrheumatoid arthritis, systemic lupus erythematosus, Kawasaki's Disease,ulcerative colitis, Crohn's Disease, ankylosing spondylitis, Behcet'ssyndrome, psoriasis, Reiter's syndrome, sarcoidosis, diabetes, multiplesclerosis, etc., or any combination thereof.

In one embodiment, the methods of the present invention may be used totreat a disease or disorder of the eye in a subject that isimmunosuppressed, while in another embodiment, in a subject that isimmunodeficient, while in another embodiment, in a subject that isimmunocompetent.

In another embodiment, the methods of the present invention may be usedto prevent or treat glaucoma. In one embodiment, glaucoma ischaracterized by increased fluid pressure in the eye, which in oneembodiment, is due to slowed fluid drainage from the eye. In oneembodiment, glaucoma may damage the optic nerve and other parts of theeye, lead to vision loss or blindness, or a combination thereof. In oneembodiment, glaucoma may refer to primary open angle glaucoma, normalpressure glaucoma, normal tension glaucoma, pigmentary glaucoma,pseudoexfoliation glaucoma, acute angle closure glaucoma, absoluteglaucoma chronic glaucoma, congenital glaucoma, juvenile glaucoma,narrow angle glaucoma, chronic open angle glaucoma, simplex glaucoma,primary congenital glaucoma, secondary glaucoma, or a combinationthereof.

In another embodiment, the methods of the present invention may be usedto prevent or treat macular degeneration. In one embodiment, maculardegeneration is characterized by damage to or breakdown of the macula,which in one embodiment, is a small area at the back of the eye. In oneembodiment, macular degeneration causes a progressive loss of centralsight, but not complete blindness. In one embodiment, maculardegeneration is of the dry type, while in another embodiment, it is ofthe wet type. In one embodiment, the dry type is characterized by thethinning and loss of function of the macula tissue. In one embodiment,the wet type is characterized by the growth of abnormal blood vesselsbehind the macula. In one embodiment, the abnormal blood vesselshemorrhage or leak, resulting in the formation of scar tissue ifuntreated. In some embodiments, the dry type of macular degeneration canturn into the wet type. In one embodiment, macular degeneration isage-related, which in one embodiment is caused by an ingrowth ofchorioidal capillaries through defects in Bruch's membrane withproliferation of fibrovascular tissue beneath the retinal pigmentepithelium.

In another embodiment, the methods of the present invention may be usedto prevent or treat retinopathy. In one embodiment, retinopathy refersto a disease of the retina, which in one embodiment is characterized byinflammation and in another embodiment, is due to blood vessel damageinside the eye. In one embodiment, retinopathy is diabetic retinopathywhich, in one embodiment, is a complication of diabetes that is causedby changes in the blood vessels of the retina. In one embodiment, bloodvessels in the retina leak blood and/or grow fragile, brush-likebranches and scar tissue, which in one embodiment, blurs or distorts theimages that the retina sends to the brain. In another embodiment,retinopathy is proliferative retinopathy, which in one embodiment, ischaracterized by the growth of new, abnormal blood vessels on thesurface of the retina (neovascularization). In one embodiment,neovascularization around the pupil increases pressure within the eye,which in one embodiment, leads to glaucoma. In another embodiment,neovascularization leads to new blood vessels with weaker walls thatbreak and bleed, or cause scar tissue to grow, which in one embodiment,pulls the retina away from the back of the eye (retinal detachment). Inone embodiment, the pathogenesis of retinopathy is related tonon-enzymatic glycation, glycoxidation, accumulation of advancedglycation end-products, free radical-mediated protein damage,up-regulation of matrix metalloproteinases, elaboration of growthfactors, secretion of adhesion molecules in the vascular endothelium, ora combination thereof.

In one embodiment, retinopathy leads to macular edema, which in oneembodiment, is swelling of the retina. In one embodiment, macular edemais characterized by retinal blood vessels that develop tiny leaks, whichin one embodiment, allow blood and fluid to seep from the retinal bloodvessels, and fatty material (called exudate) to deposit in the retina.In one embodiment, symptoms of macular edema comprise impaired orblurred vision.

In another embodiment, retinopathy refers to retinopathy of prematurity(ROP), which in one embodiment, occurs in premature babies when abnormalblood vessels and scar tissue grow over the retina. In one embodiment,retinopathy of prematurity is caused by a therapy necessary to promotethe survival of a premature infant.

In another embodiment, retinopathy refers to arterioscleroticretinopathy, which in one embodiment, is due to arteriosclerosis(hardening of the arteries). In another embodiment, retinopathy refersto hypertensive retinopathy, which in one embodiment, is due to highblood pressure. In another embodiment, retinopathy refers to solarretinopathy, while in another embodiment, it refers to drug-relatedretinopathy.

In another embodiment, the methods of the present invention may be usedto prevent or treat retinal detachment, including, inter alia,rhegmatogenous, tractional, or exudative retinal detachment, which inone embodiment, is the separation of the retina from its supportinglayers. In one embodiment, retinal detachment is associated with a tearor hole in the retina through which the internal fluids of the eye mayleak. In one embodiment, retinal detachment is caused by trauma, theaging process, severe diabetes, an inflammatory disorder,neovascularization, or retinopathy of prematurity, while in anotherembodiment, it occurs spontaneously. In one embodiment, bleeding fromsmall retinal blood vessels may cloud the vitreous during a detachment,which in one embodiment, may cause blurred and distorted images. In oneembodiment, a retinal detachment can cause severe vision loss, includingblindness.

Administration of the compounds for use in the present invention in adiversity of animal and cell models of disease invoke remarkable, andunexpected, cytoprotective effects, which are useful in the preventionand treatment of eye diseases and/or conditions.

In one embodiment of the present invention, the useful pharmacologicalproperties of the compounds for use in the present invention, some ofwhich are described hereinabove, may be applied for clinical use, anddisclosed herein as methods for the prevention or treatment of adisease. The biological basis of these methods may be readilydemonstrated by standard cellular and animal models of disease, forexample, as described in the Examples hereinbelow.

In one embodiment, the pharmacological activities of compounds for usein the present invention, including membrane stabilization,anti-inflammation, anti-oxidant action, and attenuation of chemokinelevels, may contribute to the resistance of a treated cell to diseasesof the eye. In one embodiment, cell membrane stabilization mayameliorate or prevent tissue injury arising in the course of an eyedisease. In another embodiment, anti-oxidant action may limit oxidativedamage to cell and blood components arising in the course of an eyedisease. In another embodiment, attenuation of chemokine levels mayattenuate physiological reactions to stress that arise in the course ofan eye disease.

In one embodiment of the invention, the compounds for use in the presentinvention described herein can be used to treat disease, throughamelioration or prevention, of tissue injury arising in the course ofpathological disease states by stabilizing cell membranes; limitingoxidative damage to cell and blood components; or attenuatingphysiological reactions to stress, as expressed in elevated chemokinelevels.

In one embodiment, methods of the present invention involve treating asubject by, inter alia, controlling the expression, production, andactivity of phospholipases such as PLA2; controlling the productionand/or action of lipid mediators, such as eicosanoids, plateletactivating factor (PAF) and lyso-phospholipids; amelioration of damageto cell surface glycosaminoglycans (GAG) and proteoglycans; controllingthe production of oxidants, oxygen radicals and nitric oxide; protectionof cells, tissues, and plasma lipoproteins from damaging agents, such asreactive oxygen species (ROS) and phospholipases; controlling theexpression, production, and activity of cytokines, chemokines andinterleukins; anti-oxidant therapy; anti-endotoxin therapy or anycombination thereof.

In one embodiment of the invention, the term “controlling” refers toinhibiting the production and action of the above mentioned factors inorder to maintain their activity at the normal basal level and suppresstheir activation in pathological conditions.

In one embodiment of the invention, eye disease is characterized by thepresence of damaging agents, which comprise, inter alia, phospholipases,reactive oxygen species (ROS), free radicals, lysophospholipids, fattyacids or derivatives thereof, hydrogen peroxides, phospholipids,oxidants, cationic proteins, streptolysins, proteases, hemolysins, orsialidases.

Dosages and Routes of Administration

This invention encompasses administration of compounds as describedherein or compositions comprising the same, for treating diseases of theeye.

In one embodiment, compositions of this invention are pharmaceuticallyacceptable. In one embodiment, the term “pharmaceutically acceptable”refers to any formulation which is safe, and provides the appropriatedelivery for the desired route of administration of an effective amountof at least one compound for use in the present invention. This termrefers to the use of buffered formulations as well, wherein the pH ismaintained at a particular desired value, ranging from pH 4.0 to pH 9.0,in accordance with the stability of the compounds and route ofadministration.

In some embodiments, any of the compositions of this invention willcomprise a lipid conjugate, in any form or embodiment as describedherein. In some embodiments, any of the compositions of this inventionwill consist of a lipid conjugate, in any form or embodiment asdescribed herein. In some embodiments, of the compositions of thisinvention will consist essentially of a lipid conjugate, in any form orembodiment as described herein. In some embodiments, the term “comprise”refers to the inclusion of the indicated active agent, such as theCompounds I-C, as well as inclusion of other active agents, andpharmaceutically acceptable carriers, excipients, emollients,stabilizers, etc., as are known in the pharmaceutical industry. In someembodiments, the term “consisting essentially of” refers to acomposition, whose only active ingredient is the indicated activeingredient, however, other compounds may be included which are forstabilizing, preserving, etc. the formulation, but are not involveddirectly in the therapeutic effect of the indicated active ingredient.In some embodiments, the term “consisting essentially of” may refer tocomponents which facilitate the release of the active ingredient. Insome embodiments, the term “consisting” refers to a composition, whichcontains the active ingredient and a pharmaceutically acceptable carrieror excipient.

In one embodiment, a compound used in the methods of this invention maybe administered alone or within a composition. In another embodiment,compositions comprising compounds for use in the present invention inadmixture with conventional excipients, i.e. pharmaceutically acceptableorganic or inorganic carrier substances suitable for parenteral, enteral(e.g. oral) or topical application which do not deleteriously react withthe active compounds may be used. In one embodiment, suitablepharmaceutically acceptable carriers include but are not limited towater, salt solutions, alcohols, gum arabic, vegetable oils, benzylalcohols, polyethylene glycols, gelatine, carbohydrates such as lactose,amylose or starch, magnesium stearate, talc, silicic acid, viscousparaffin, white paraffin, glycerol, alginates, hyaluronic acid,collagen, perfume oil, fatty acid monoglycerides and diglycerides,pentaerythritol fatty acid esters, hydroxy methylcellulose, polyvinylpyrrolidone, etc. In another embodiment, the pharmaceutical preparationscan be sterilized and if desired mixed with auxiliary agents, e.g.lubricants, preservatives, stabilizers, wetting agents, emulsifiers,salts for influencing osmotic pressure, buffers, coloring, flavoringand/or aromatic substances and the like which do not deleteriously reactwith the active compounds. In another embodiment, they can also becombined where desired with other active agents, e.g. vitamins.

In one embodiment, the therapeutic compositions of the instant inventioncomprise a compound of the instant invention and additional compoundseffective in preventing or treating eye disease. In one embodiment, theadditional compounds comprise anti-inflammatory compositions, which inone embodiment are non-steroidal anti-inflammatory medications,antihistamines, antibiotics, corticosteroids, cromolyn sodium (sodiumcromoglicate), mast-cell stabilizers, artificial tears, lubricants, or acombination thereof. In one embodiment, antibiotics comprisechloramphenicol, fusidic acid, tetracycline, erythromycin, gentamycin,or a combination thereof. In another embodiment, an additional compoundis vitamin A.

In one embodiment, the therapeutic compositions of the instant inventionare administered with other treatments that relieve symptoms. In oneembodiment, other treatments comprise application of cold compresses,while in another embodiment, warm compresses.

In one embodiment, the route of administration may be parenteral,enteral, or a combination thereof. In another embodiment, the route maybe intra-ocular, topical, transdermal, intradermal, subcutaneous,intraperitoneal, intravenous, intra-arterial, vaginal, rectal,intratumoral, parcanceral, transmucosal, intramuscular, intravascular,intraventricular, intracranial, inhalation, nasal aspiration (spray),sublingual, oral, aerosol or suppository or a combination thereof. Inone embodiment, the dosage regimen will be determined by skilledclinicians, based on factors such as exact nature of the condition beingtreated, the severity of the condition, the age and general physicalcondition of the patient, etc.

For intra-ocular application, eye drops, ointments, lotions, creams, orcoated eye patches may be used in one embodiment. In another embodiment,intra-ocular application may comprise the use of contact lens comprisingthe compounds of the instant invention.

In one embodiment, intra-ocular application is used to treat an eyecondition or disease. In another embodiment, intra-ocular injection isused to treat an eye condition or disease. In one embodiment, compoundsmay be administered intravitreally, in another embodiment, subretinally,while in another embodiment, intra-retinally, while in anotherembodiment, periocularly. In one embodiment, compounds may beadministered intracamerally into the anterior chamber or vitreous, via adepot attached to the intraocular lens implant inserted during surgery,or via a depot placed in the eye sutured in the anterior chamber orvitreous.

For parenteral application, particularly suitable are injectable,sterile solutions, preferably oily or aqueous solutions, as well assuspensions, emulsions, or implants, including suppositories and enemas.Ampoules are convenient unit dosages. Such a suppository may compriseany agent described herein.

For application by inhalation, solutions or suspensions of the compoundsmixed and aerosolized or nebulized in the presence of the appropriatecarrier suitable. Such an aerosol may comprise any agent describedherein and, in one embodiment, may be used to treat diseases orconditions caused by airborne pathogens, which may in one embodiment,cause sinusitis or upper respiratory infections, in addition to eyediseases.

For topical application, particularly in the area around the eye, anadmixture of the compounds with conventional creams, lotions, or delayedrelease patches is acceptable. Such a cream or lotion may comprise anyagent described herein, and, in one embodiment, may be used to treat aneye disease.

For enteral application, particularly suitable are tablets, dragees,liquids, drops, or capsules. A syrup, elixir, or the like can be usedwhen a sweetened vehicle is employed.

Sustained or directed release compositions can be formulated, e.g.liposomes or those wherein the active compound is protected withdifferentially degradable coatings, e.g. by microencapsulation, multiplecoatings, etc. It is also possible to freeze-dry the new compounds anduse the lyophilisates obtained, for example, for the preparation ofproducts for injection.

Thus, in one embodiment, the route of administration may be directed toan organ or system that is affected by an eye disease. For example,compounds may be administered in intra-ocular form to treat an eyedisease. In another embodiment, the route of administration may bedirected to a different organ or system than the one that is affected byan eye disease. For example, compounds may be administered parenterallyto treat an eye disease. Thus, the present invention provides for theuse of compounds of the instant invention in various dosage formssuitable for administration using any of the routes listed hereinabove.

In general, the doses utilized for the above described purposes willvary, but will be in an effective amount to exert the desired effect. Asused herein, the term “pharmaceutically effective amount” refers to anamount of a compound of formulae A and I-LXXXVII as describedhereinbelow, which will produce the desired alleviation in symptoms orother desired phenotype in a patient. The doses utilized for any of theabove-described purposes will generally be from 1 to about 1000milligrams per kilogram of body weight (mg/kg), administered one to fourtimes per day, or by continuous IV infusion. When the compositions aredosed topically or intraocularly, they will generally be in aconcentration range of from 0.1 to about 10% w/v, administered 1-4 timesper day.

In one embodiment of the invention, the concentrations of the compoundswill depend on various factors, including the nature of the condition tobe treated, the condition of the patient, the route of administrationand the individual tolerability of the compositions.

It will be appreciated that the actual preferred amounts of activecompound in a specific case will vary according to the specific compoundbeing utilized, the particular compositions formulated, the mode ofapplication, and the particular conditions and organism being treated.Dosages for a given host can be determined using conventionalconsiderations, e.g. by customary comparison of the differentialactivities of the subject compounds and of a known agent, e.g. by meansof an appropriate, conventional pharmacological protocol.

In one embodiment, the compounds of the invention may be administeredacutely for acute treatment of temporary conditions, or may beadministered chronically, especially in the case of progressive,recurrent, or degenerative disease. In one embodiment, one or morecompounds of the invention may be administered simultaneously, or inanother embodiment, they may be administered in a staggered fashion. Inone embodiment, the staggered fashion may be dictated by the stage orphase of the disease.

In one embodiment, the present invention offers methods for thetreatment of disease based upon administration of lipids covalentlyconjugated through their polar head group to a physiologicallyacceptable chemical moiety, which may be of high or low molecularweight.

The present invention has been illustrated in terms of the anti-diseaseactivity of compounds for use in the present invention and methods oftheir use as pharmaceutical compositions in the treatment of disease.The following sections present some examples of the therapeuticcompounds for use in the present invention and their chemicalpreparation.

Compounds

In one embodiment, the compounds for use in the present invention or forthe compositions of the present invention comprise a lipid orphospholipid moiety bound to a physiologically acceptable monomer,dimer, oligomer, or polymer. In one embodiment, the physiologicallyacceptable monomer, dimer, oligomer, or polymer is salicylate, salicylicacid, aspirin, a monosaccharide, lactobionic acid, maltose, an aminoacid, glycine, carboxylic acid, acetic acid, butyric acid, dicarboxylicacid, glutaric acid, succinic acid, fatty acid, dodecanoic acid,didodecanoic acid, bile acid, cholic acid, cholesterylhemmisuccinate, adipeptide, a disaccharide, a trisaccharide, an oligosaccharide, apolysaccharide, a hetero-polysaccharide, a homo-polysaccharide, apolypyranose, an oligopeptide, or a di- or trisaccharide monomer unit ofheparin, heparan sulfate, keratin, keratan sulfate, chondroitin,chondroitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatansulfate, dextran, or hyaluronic acid, a glycosaminoglycan, polygeline(‘haemaccel’), alginate, hydroxyethyl starch (hetastarch), polyethyleneglycol, polycarboxylated polyethylene glycol, chondroitin-6-sulfate,chondroitin-4-sulfate, keratin, keratin sulfate, heparan sulfate,dermatin, dermatan sulfate, carboxymethylcellulose, heparin, dextran, orhyaluronic acid.

In one embodiment, examples of polymers which can be employed as theconjugated moiety for producing compounds for use in the presentinvention for use in the methods of this invention may bephysiologically acceptable polymers, including water-dispersible or-soluble polymers of various molecular weights and diverse chemicaltypes, mainly natural and synthetic polymers, such asglycosaminoglycans, hyaluronic acids, heparin, heparin sulfates,chondroitin sulfates, chondroitin-6-sulfates, chondroitin-4-sulfates,keratins, keratin sulfates, dermatins, dermatan sulfates, dextrans,plasma expanders, including polygeline (“Haemaccel”, degraded gelatinpolypeptide cross-linked via urea bridges, produced by “Behring”),“hydroxyethylstarch” (Hetastarch, HES) and extrans, food and drugadditives, soluble cellulose derivatives (e.g. methylcellulose,carboxymethylcellulose), polyaminoacids, hydrocarbon polymers (e.g.polyethylene), polystyrenes, polyesters, polyamides, polyethylene oxides(e.g. polyethyleneglycols, polycarboxyethyleneglycols, polycarboxylatedpolyethyleneglycols), polyvinnylpyrrolidones, polysaccharides,polypyranoses, alginates, assimilable gums (e.g. xanthan gum), peptides,injectable blood proteins (e.g. serum albumin), cyclodextrin, andderivatives thereof.

In one embodiment, examples of monomers, dimers, and oligomers which canbe employed as the conjugated moiety for producing compounds for use inthe present invention for use in the methods of the invention may bemono- or disaccharides, trisaccharides, oligopeptides, carboxylic acids,dicarboxylic acids, fatty acids, dicarboxylic fatty acids, salicylates,slicyclic acids, acetyl salicylic acids, aspirins, lactobionic acids,maltoses, amino acids, glycines, glutaric acids, succinic acids,dodecanoic acids, didodecanoic acids, bile acids, cholic acids,cholesterylhemisuccinates, and di- and trisaccharide unit monomers ofpolysaccharides, polypyranoses, and/or glycosaminoglycans includingheparins, heparan sulfates, hyaluronic acids, chondroitins, chondroitinsulfates, chondroitin-6-sulfates, chondroitin-4-sulfates, dermatins,dermatan sulfates, keratins, keratan sulfates, or dextrans.

In one embodiment, the lipid compounds for use in the present inventionare described by the general formula:

[phosphatidylethanolamine-Y]n-X

[phosphatidylserine-Y]n-X

[phosphatidylcholine-Y]n-X

[phosphatidylinositol-Y]n-X

[phosphatidylglycerol-Y]n-X

[phosphatidic acid-Y]n-X

[lyso-phospholipid-Y]n-X

[diacyl-glycerol-Y]n-X

[monoacyl-glycerol-Y]n-X

[sphingomyelin-Y]n-X

[sphingosine-Y]n-X

[ceramide-Y]n-X

wherein

-   -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms; and    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer; and    -   n is the number of lipid molecules bound to a molecule of X,        wherein n is a number from 1 to 1000. In another embodiment, n        is a number from 2 to 1000.

In one embodiment, the invention provides low-molecular weightcompounds, previously undisclosed and unknown to possess pharmacologicalactivity, of the general formula described hereinabove. In anotherembodiment, wherein the general formula described hereinabove describeslow-molecular weight compounds, X is a mono- or disaccharide,carboxylated disaccharide, mono- or dicarboxylic acids, a salicylate,salicylic acid, aspirin, lactobionic acid, maltose, an amino acid,glycine, acetic acid, butyric acid, dicarboxylic acid, glutaric acid,succinic acid, fatty acid, dodecanoic acid, didodecanoic acid, bileacid, cholic acid, cholesterylhemmisuccinate, a di- or tripeptide, anoligopeptide, a trisacharide, or a di- or trisaccharide monomer unit ofheparin, heparan sulfate, keratin, keratan sulfate, chondroitin,chondroitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatansulfate, dextran, or hyaluronic acid.

In one embodiment of this invention, X is any of the physiologicallyacceptable monomer, dimer, oligomer, or polymer, as described herein. Inone embodiment, X is conjugated to the lipid, phospholipid, or spacervia an ester bond. In another embodiment, X is conjugated to the lipid,phospholipid, or spacer via an amide bond.

As defined by the structural formulae provided herein for the compoundsfor use in the present invention, these compounds may contain betweenone to one thousand lipid moieties bound to a single physiologicallyacceptable polymer molecule. In one embodiment of this invention, n is anumber from 1 to 1000. In another embodiment, n is a number from 1 to500. In another embodiment, n is a number from 1 to 100. In anotherembodiment, n is a number from 1 to 50. In another embodiment, n is anumber from 1 to 25. In another embodiment, n is a number from 1 to 10.In another embodiment, n is a number from 1-5. In another embodiment, nis a number from 1 to 4. In another embodiment, n is a number from 1 to3, In another embodiment, n is a number from 1 to 2. In anotherembodiment, n is a number from 2 to 1000. In another embodiment, n is anumber from 2 to 100. In another embodiment, n is a number from 2 to200. In another embodiment, n is a number from 2 to 50. In anotherembodiment, n is a number from 2 to 25. In another embodiment, n is anumber from 2-10. In another embodiment, n is a number from 2 to 5. Inanother embodiment, n is a number from 2 to 4, In another embodiment, nis a number from 2 to 3. In another embodiment, n is a number from 3 to300. In another embodiment, n is a number from 10 to 400. In anotherembodiment, n is a number from 50 to 500. In another embodiment, n is anumber from 100 to 300. In another embodiment, n is a number from 300 to500. In another embodiment, n is a number from 500 to 800. In anotherembodiment, n is a number from 500 to 1000.

In one embodiment of the invention, when the conjugated moiety is apolymer, the ratio of lipid moieties covalently bound may range from oneto one thousand lipid residues per polymer molecule, depending upon thenature of the polymer and the reaction conditions employed. For example,the relative quantities of the starting materials, or the extent of thereaction time, may be modified in order to obtain products with eitherhigh or low ratios of lipid residues per polymer, as desired.

In one embodiment, the set of compounds comprisingphosphatidylethanolamine covalently bound to a physiologicallyacceptable monomer, dimmer, oligomer, or polymer, is referred to hereinas the PE-conjugates. In one embodiment, the phosphatidylethanolaminemoiety is dipalmitoyl phosphatidylethanolamine. In another embodiment,the phosphatidylethanolamine moiety is dimyristoylphosphatidylethanolamine. In another embodiment, related derivatives, inwhich either phosphatidylserine, phosphatidylcholine,phosphatidylinositol, phosphatidic acid or phosphatidylglycerol areemployed in lieu of phosphatidylethanolamine as the lipid moiety provideequivalent therapeutic results, based upon the biological experimentsdescribed below for the compounds for use in the present invention andthe structural similarities shared by these compounds.

In another embodiment, the lipid or phospholipid moiety is phosphatidicacid, an acyl glycerol, monoacylglycerol, diacylglycerol,triacylglycerol, sphingosine, sphingomyelin, chondroitin-4-sulfate,chondroitin-6-sulfate, ceramide, phosphatidylethanolamine,phosphatidylserine, phosphatidylcholine, phosphatidylinositol, orphosphatidylglycerol, or an ether or alkyl phospholipid derivativethereof.

In one embodiment, derivatives relevant to this invention are compoundswherein at least one of the fatty acid groups of the lipid moieties atposition C1 or C2 of the glycerol backbone are substituted by a longchain alkyl group attached by amide, ether or alkyl bonds, rather thanester linkages.

In the methods, according to embodiments of the invention, the compoundsfor use in the present invention administered to the subject arecomprised from at least one lipid moiety covalently bound through anatom of the polar head group to a monomeric or polymeric moiety(referred to herein as the conjugated moiety) of either low or highmolecular weight. When desired, an optional bridging moiety can be usedto link the compounds for use in the present invention moiety to themonomer or polymeric moiety. The conjugated moiety may be a lowmolecular weight carboxylic acid, dicarboxylic acid, fatty acid,dicarboxylic fatty acid, acetyl salicylic acid, cholic acid,cholesterylhemisuccinate, or mono- or di-saccharide, an amino acid ordipeptide, an oligopeptide, a glycoprotein mixture, a di- ortrisaccharide monomer unit of a glycosaminoglycan such as a repeatingunit of heparin, heparan sulfate, hyaluronic acid, chondroitin-sulfate,dermatan, keratan sulfate, or a higher molecular weight peptide oroligopeptide, a polysaccharide, a hetero-polysaccharide, ahomo-polysaccharide, a polypyranose, polyglycan, protein,glycosaminoglycan, or a glycoprotein mixture. The composition of somephospholipid-conjugates of high molecular weight, and associatedanalogues, are the subject of U.S. Pat. No. 5,064,817, which isincorporated herein in its entirety by reference.

In one embodiment, the term “moiety” means a chemical entity otherwisecorresponding to a chemical compound, which has a valence satisfied by acovalent bond.

In some cases, according to embodiments of the invention, the monomer orpolymer chosen for preparation of the compound may in itself have selectbiological properties. For example, both heparin and hyaluronic acid arematerials with known physiological functions. In the present invention,however, the compounds for use in the present invention formed fromthese substances as starting materials display a new and wider set ofpharmaceutical activities than would be predicted from administration ofeither heparin or hyaluronic acid which have not been bound by covalentlinkage to a phospholipid. In some embodiments, phosphatidylethanolamine(PE) linked to hyaluronic acid (Compound XXII), to heparin (CompoundXXIV), to chondroitin sulfate A (Compound XXV), tocarboxymethylcellulose (Compound XXVI), to Polygeline (haemaccel)(Compound XXVII), to alginate (Compound LI), or to hydroxyethylstarch(Compound XXVIII), are useful for methods and in compositions as hereindescribed but perform unexpectedly in terms of potency and range ofuseful pharmaceutical activity compared to the free conjugates. Thus,the combination of a phospholipid such as phosphatidylethanolamine, orrelated phospholipids which differ with regard to the polar head group,such as phosphatidylserine (PS), phosphatidylcholine (PC),phosphatidylinositol (PI), and phosphatidylglycerol (PG), results in theformation of a compound which has novel pharmacological properties whencompared to the starting materials alone. In one embodiment, suchproperties may include: greater lubrication, greater local persistence,greater anti-inflammatory properties, greater antioxidant activity, or acombination thereof.

The biologically active compounds for use in the present inventiondescribed herein can have a wide range of molecular weights, e.g. above50,000 (up to a few hundred thousands) when it is desirable to retainthe lipid conjugate in the vascular system and below 50,000 whentargeting to extravascular systems is desirable. The sole limitation onthe molecular weight and the chemical structure of the conjugated moietyis that it does not result in a compound devoid of the desiredbiological activity, or lead to chemical or physiological instability tothe extent that the Compound is rendered useless as a drug in the methodof use described herein.

In one embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline,        phosphate, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000;    -   wherein any bond between L, Z, Y and X is either an amide or an        esteric bond.

In one embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline,        phosphate, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 2 to 1000;    -   wherein any bond between L, Z, Y and X is either an amide or an        esteric bond.

In one embodiment, L is phosphatidyl, Z is ethanolamine, wherein L and Zare chemically bonded resulting in phosphatidylethanolamine, Y isnothing, and X is carboxymethylcellulose. In another embodiment, L isphosphatidyl, Z is ethanolamine, wherein L and Z are chemically bondedresulting in phosphatidylethanolamine, Y is nothing, and X is aglycosaminoglycan. In one embodiment, the phosphatidylethanolaminemoiety is dipalmitoyl phosphatidylethanolamine. In another embodiment,the phosphatidylethanolamine moiety is dimyristoylphosphatidylethanolamine.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (I):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms; and    -   X is either a physiologically acceptable monomer, dimer,        oligomer or a physiologically acceptable polymer; and    -   n is a number from 1 to 1,000;    -   wherein if Y is nothing the phosphatidylethanolamine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylethanolamine via an amide bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (I):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms; and    -   X is either a physiologically acceptable monomer, dimer,        oligomer or a physiologically acceptable polymer; and    -   n is a number from 2 to 1,000;    -   wherein if Y is nothing the phosphatidylethanolamine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylethanolamine via an amide bond.

In one embodiment, compounds for use in the methods of the inventioncomprise one of the following as the conjugated moiety X: acetate,butyrate, glutarate, succinate, dodecanoate, didodecanoate, maltose,lactobionic acid, dextran, alginate, aspirin, cholate,cholesterylhemisuccinate, carboxymethyl-cellulose, heparin, hyaluronicacid, chondroitin sulfate, polygeline (haemaccel), hydroxyethylstarch(Hetastarch, HES) polyethyleneglycol, polycarboxylated polyethyleneglycol, a glycosaminoglycan, a polysaccharide, a hetero-polysaccharide,a homo-polysaccharide, or a polypyranose. The polymers used as startingmaterial to prepare the PE-conjugates may vary in molecular weight from1 to 2,000 kDa.

Examples of phosphatidylethanolamine (PE) moieties are analogues of thephospholipid in which the chain length of the two fatty acid groupsattached to the glycerol backbone of the phospholipid varies from 2-30carbon atoms length, and in which these fatty acids chains containsaturated and/or unsaturated carbon atoms. In lieu of fatty acid chains,alkyl chains attached directly or via an ether linkage to the glycerolbackbone of the phospholipid are included as analogues of PE. In oneembodiment, the PE moiety is dipalmitoyl-phosphatidyl-ethanolamine. Inanother embodiment, the PE moiety isdimyristoyl-phosphatidyl-ethanolamine.

Phosphatidyl-ethanolamine and its analogues may be from various sources,including natural, synthetic, and semi-synthetic derivatives and theirisomers.

Phospholipids which can be employed in lieu of the PE moiety areN-methyl-PE derivatives and their analogues, linked through the aminogroup of the N-methyl-PE by a covalent bond; N,N-dimethyl-PE derivativesand their analogues linked through the amino group of theN,N-dimethyl-PE by a covalent bond, phosphatidylserine (PS) and itsanalogues, such as palmitoyl-stearoyl-PS, natural PS from varioussources, semi-synthetic PSs, synthetic, natural and artifactual PSs andtheir isomers. Other phospholipids useful as conjugated moieties in thisinvention are phosphatidylcholine (PC), phosphatidylinositol (PI),phosphatidic acid and phosphoatidylglycerol (PG), as well as derivativesthereof comprising either phospholipids, lysophospholipids, phosphatidicacid, sphingomyelins, lysosphingomyelins, ceramide, and sphingosine.

For PE-conjugates and PS-conjugates, the phospholipid is linked to theconjugated monomer or polymer moiety through the nitrogen atom of thephospholipid polar head group, either directly or via a spacer group.For PC, PI, and PG conjugates, the phospholipid is linked to theconjugated monomer or polymer moiety through either the nitrogen or oneof the oxygen atoms of the polar head group, either directly or via aspacer group.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (II):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein if Y is nothing, the phosphatidylserine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylserine via an amide bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (II):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein if Y is nothing, the phosphatidylserine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylserine via an amide bond.

In one embodiment, the phosphatidylserine may be bonded to Y, or to X ifY is nothing, via the COO⁻ moiety of the phosphatidylserine.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (III):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phosphatidyl, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IV):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (III):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phosphatidyl, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IV):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IV):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (V):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (V):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In one embodiment of the invention, phosphatidylcholine (PC),phosphatidylinositol (PI), phosphatidic acid (PA), wherein Z is nothing,and phosphatidylglycerol (PG) conjugates are herein defined as compoundsof the general formula (III).

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VIII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VIII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IXa):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IXa):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IXb):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IXb):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (X):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the ceramide phosphoryl, Z, Y and X is        either an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (X):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;

wherein any bond between the ceramide phosphoryl, Z, Y and X is eitheran amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XI):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein if Y is nothing the sphingosyl is directly linked to X        via an amide bond and if Y is a spacer, the spacer is directly        linked to X and to the sphingosyl via an amide bond and to X via        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XI):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein if Y is nothing the sphingosyl is directly linked to X        via an amide bond and if Y is a spacer, the spacer is directly        linked to X and to the sphingosyl via an amide bond and to X via        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the ceramide, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the ceramide, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIII):

(XIII)

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the diglyceryl, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the diglyceryl, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIV):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the glycerolipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIV):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the glycerolipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XV):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the glycerolipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XV):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the glycerolipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVII):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVII):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVIII):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVIII):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XXI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XXI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a glycosaminoglycan; and    -   n is a number from 2 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

For any or all of the compounds represented by the structures of thegeneral formulae (A), (I), (II), (III), (IV), (V), (VI), (VII), (VIII),(IX), (IXa), (IXb), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI),(XVII), (XVIII), (XIX), (XX), (XXI), and (XXII) hereinabove: In oneembodiment, X is a glycosaminoglycan. According to this aspect and inone embodiment, the glycosaminoglycan may be, inter alia, hyaluronicacid, heparin, heparan sulfate, chondroitin sulfate, keratin, keratansulfate, dermatan sulfate or a derivative thereof. In anotherembodiment, X is not a glycosaminoglycan. In another embodiment, X is apolysaccharide, which in one embodiment is a hetero-polysaccharide, andin another embodiment, is a homo-polysaccharide. In another embodiment,X is a polypyranose.

In another embodiment, the glycosaminoglycan is a polymer ofdisaccharide units. In another embodiment, the number of thedisaccharide units in the polymer is m. In another embodiment, m is anumber from 2-10,000. In another embodiment, m is a number from 2-500.In another embodiment, m is a number from 2-1000. In another embodiment,m is a number from 50-500. In another embodiment, m is a number from2-2000. In another embodiment, m is a number from 500-2000. In anotherembodiment, m is a number from 1000-2000. In another embodiment, m is anumber from 2000-5000. In another embodiment, m is a number from3000-7000. In another embodiment, m is a number from 5000-10,000. Inanother embodiment, a disaccharide unit of a glycosaminoglycan may bebound to one lipid or phospholipid moiety. In another embodiment, eachdisaccharide unit of the glycosaminoglycan may be bound to zero or onelipid or phospholipid moieties. In another embodiment, the lipid orphospholipid moieties are bound to the —COOH group of the disaccharideunit. In another embodiment, the bond between the lipid or phospholipidmoiety and the disaccharide unit is an amide bond.

In one embodiment, the invention encompasses compounds, compositions andpreparations, comprising a phospholipid-GAG conjugate, whereby the molarratio in the compounds, compositions and preparations between thephospholipid and the GAG is in the range of between 1.5:1 to 20:1, or inanother embodiments, 2:1 to 10:1, or in another embodiment, 3:1 to 7:1,or in another embodiment, 1.5:1 to 7:1. In another embodiment, the molarratio between the phospholipid and the GAG is in the range of between2:1 to 10:1. In another embodiment, the molar ratio between thephospholipid and the GAG is in the range of between 2:1 to 5:1. Inanother embodiment, the molar ration between the phospholipid and theGAG is 2:1. In another embodiment, the molar ration between thephospholipid and the GAG is 3:1. In another embodiment, the molar rationbetween the phospholipid and the GAG is 5:1. In another embodiment, themolar ration between the phospholipid and the GAG is 10:1. In anotherembodiment, the molar ration between the phospholipid and the GAG is20:1.

In one embodiment, the compound of the present invention comprises aglycosaminoglycan (GAG) with a molecular weight in the range of between30-100 kD. In another embodiment, the GAG has a molecular weight in therange of between 30-80 kD. In another embodiment, the GAG has amolecular weight in the range of between 30-50 kD. In anotherembodiment, the GAG has a molecular weight in the range of between 20-80kD. In another embodiment, the GAG has a molecular weight in the rangeof between 20-50 kD.

In another embodiment, the chondroitin sulfate may be, inter alia,chondroitin-6-sulfate, chondroitin-4-sulfate or a derivative thereof.

In one embodiment of the invention, Y is nothing. Non-limiting examplesof suitable divalent groups forming the optional bridging group (whichin one embodiment, is referred to as a spacer) Y, according toembodiments of the invention, are straight or branched chain alkylene,e.g. of 2 or more, preferably 4 to 30 carbon atoms, —CO-alkylene-CO,—NH-alkylene-NH—, —CO-alkylene-NH—, —NH-alkylene-NH, CO-alkylene-NH—, anamino acid, cycloalkylene, wherein alkylene in each instance, isstraight or branched chain and contains 2 or more, preferably 2 to 30atoms in the chain, —(—O—CH(CH₃)CH₂)_(x) wherein x is an integer of 1 ormore.

According to embodiments of the invention, in addition to thetraditional phospholipid structure, related derivatives for use in thisinvention are phospholipids modified at the C1 or C2 position to containan amine, ether or alkyl bond instead of an ester bond. In oneembodiment of the invention, the alkyl phospholipid derivatives andether phospholipid derivatives are exemplified herein.

In one embodiment of the invention, the sugar rings of theglycosaminoglycan are intact. In another embodiment, intact refers toclosed. In another embodiment, intact refers to natural. In anotherembodiment, intact refers to unbroken.

In one embodiment of the invention, the structure of the lipid orphospholipid in any compound according to the invention is intact. Inanother embodiment, the natural structure of the lipid or phospholipidsin any compound according to the invention is maintained.

In one embodiment, the compounds for use in the present invention arebiodegradable.

In one embodiment, the compound according to the invention isphosphatidylethanolamine bound to aspirin. In one embodiment, thecompound according to the invention is phosphatidylethanolamine bound toglutarate.

In some embodiments, the compounds for use are as listed in Table 1below.

TABLE 1 Phospholipid Spacer Polymer (m.w.) Compound PE None Hyaluronicacid (2-2000 kDa) XXII Dimyristoyl-PE None Hyaluronic acid XXIII PE NoneHeparin (0.5-110 kDa) XXIV PE None Chondroitin sulfate A XXV PE NoneCarboxymethylcellulose (20-500 kDa) XXVI PE Dicarboxylic acid + DiaminePolygeline (haemaccel) (4-40 kDa) XXVII PE None HydroxyethylstarchXXVIII PE Dicarboxylic acid + Diamine Dextran (1-2,000 kDa) XXIX PE NoneAspirin XXX PE Carboxyl amino group Hyaluronic acid (2-2000 kDa) XXXI PEDicarboxyl group Hyaluronic acid (2-2000 kDa) XXXII PE Dipalmitoic acidHyaluronic acid (2-2000 kDa) XXXIII PE Carboxyl amino group Heparin(0.5-110 kDa) XXXIV PE Dicarboxyl group Heparin (0.5-110 kDa) XXXV PECarboxyl amino group Chondroitin sulfate A XXXVI PE Dicarboxyl groupChondroitin sulfate A XXXVII PE Carboxyl amino groupCarboxymethylcellulose (20-500 kDa) XXXVIII PE Dicarboxyl groupCarboxymethylcellulose (20-500 kDa) XXXIX PE None Polygeline (haemaccel)(4-40 kDa) XL PE Carboxyl amino group Polygeline (haemaccel) (4-40 kDa)XLI PE Dicarboxyl group Polygeline (haemaccel) (4-40 kDa) XLII PECarboxyl amino group Hydroxyethylstarch XLIII PE Dicarboxyl groupHydroxyethylstarch XLIV PE None Dextran (1-2,000 kDa) XLV PE Carboxylamino group Dextran (1-2,000 kDa) XLVI PE Dicarboxyl group Dextran(1-2,000 kDa) XLVII PE Carboxyl amino group Aspirin XLVIII PE Dicarboxylgroup Aspirin XLIX PE None Albumin L PE None Alginate (2-2000 kDa) LI PENone Polyaminoacid LII PE None Polyethylene glycol LIII PE NoneLactobionic acid LIV PE None Acetylsalicylate LV PE NoneCholesteryl-hemmisuccinate LVI PE None Maltose LVII PE None Cholic acidLVIII PE None Chondroitin sulfates LIX PE None Polycarboxylatedpolyethylene glycol LX Dipalmitoyl-PE None Hyaluronic acid LXIDipalmitoyl-PE None Heparin LXII Dipalmitoyl-PE None Chondroitin sulfateA LXIII Dipalmitoyl-PE None Carboxymethylcellulose LXIV Dipalmitoyl-PENone Polygeline (haemaccel) LXV Dipalmitoyl-PE None HydroxyethylstarchLXVI Dipalmitoyl-PE None Dextran LXVII Dipalmitoyl-PE None AspirinLXVIII Dimyristoyl-PE None Heparin LXVIX Dimyristoyl-PE None Chondroitinsulfate A LXX Dimyristoyl-PE None Carboxymethylcellulose LXXIDimyristoyl-PE None Polygeline (haemaccel) LXXII Dimyristoyl-PE NoneHydroxyethylstarch LXXIII Dimyristoyl-PE None Dextran LXXIVDimyristoyl-PE None Aspirin LXXV PS None Hyaluronic acid LXXVI PS NoneHeparin LXXVII PS None Polygeline (haemaccel) LXXVIII PC None Hyaluronicacid LXXIX PC None Heparin LXXX PC None Polygeline (haemaccel) LXXXI PINone Hyaluronic acid LXXXII PI None Heparin LXXXIII PI None Polygeline(haemaccel) LXXXIV PG None Hyaluronic acid LXXXV PG None Heparin LXXXVIPG None Polygeline (haemaccel) LXXXVII PE None Glutaryl LXXXVIIIDipalmitoyl-PE None Alginate LXXXIX Dimyristoyl-PE None Alginate XC PSNone Alginate XCI PC None Alginate XCII PI None Alginate XCIII PG NoneAlginate XCIV PS None Hydroxyethylstarch XCV PC None HydroxyethylstarchXCVI PI None Hydroxyethylstarch XCVII PG None Hydroxyethylstarch XCVIIIPE —CO—(CH₂)₃—CO—NH—(CH₂)₆— Hydroxyethylstarch XCIX PE —CO—CH₂—Carboxymethylcellulose C

In one embodiment of the invention, the compounds for use in the presentinvention are any one or more of Compounds I-C. In another embodiment,the invention provides a composition comprising any combination of anyof the compounds of the invention or the use of any combination of anyof the compounds of the invention. In another embodiment, the inventionprovides a composition comprising Compounds XCIX, C, or a combinationthereof and uses thereof. In another embodiment, the invention providesa composition comprising Compounds LXV, LXVI, LXXI, LXXII, LXXIII,LXXXIX, XC, or a combination thereof and uses thereof. In anotherembodiment, the compounds for use in the present invention are CompoundXXII, Compound XXIII, Compound XXIV, Compound XXV, Compound XXVI,Compound XXVII, Compound XXVIII, Compound XXIX, Compound XXX, CompoundLI, or pharmaceutically acceptable salts thereof, in combination with aphysiologically acceptable carrier or solvent. According to embodimentsof the invention, these polymers, when chosen as the conjugated moiety,may vary in molecular weights from 200 to 2,000,000 Daltons. In oneembodiment of the invention, the molecular weight of the polymer asreferred to herein is from 200 to 1000 Daltons. In another embodiment,the molecular weight of the polymer as referred to herein is from 200 to1000 Daltons. In another embodiment, the molecular weight of the polymeras referred to herein is from 1000 to 5000 Daltons. In anotherembodiment, the molecular weight of the polymer as referred to herein isfrom 5000 to 10,000 Daltons. In another embodiment, the molecular weightof the polymer as referred to herein is from 10,000 to 20,000 Daltons.In another embodiment, the molecular weight of the polymer as referredto herein is from 10,000 to 50,000 Daltons. In another embodiment, themolecular weight of the polymer as referred to herein is from 20,000 to70,000 Daltons. In another embodiment, the molecular weight of thepolymer as referred to herein is from 50,000 to 100,000 Daltons. Inanother embodiment, the molecular weight of the polymer as referred toherein is from 100,000 to 200,000 Daltons. In another embodiment, themolecular weight of the polymer as referred to herein is from 200,000 to500,000 Daltons. In another embodiment, the molecular weight of thepolymer as referred to herein is from 200,000 to 1,000,000 Daltons. Inanother embodiment, the molecular weight of the polymer as referred toherein is from 500,000 to 1,000,000 Daltons. In another embodiment, themolecular weight of the polymer as referred to herein is from 1,000,000to 2,000,000 Daltons. Various molecular weight species have been shownto have the desired biological efficacy.

In one embodiment, AlgPE has a molecular weight of approximately 120 kD,CSAPE has a molecular weight of approximately 100 kD, HemPE has amolecular weight of approximately 75 kD, HesDMPE has a molecular weightof approximately 90 kD, CMPE has a molecular weight of approximately 75kD, or a combination thereof. In one embodiment, “approximately” refersto up to 5%, 10%, 15%, 20%, or 25% of the value. In another embodiment,“approximately” refers to 5-25%, 5-15%. 10-25%, 10-20%, 15-25% of thevalue.

In one embodiment of this invention, low molecular weight compounds foruse in the present invention are defined hereinabove as the compounds offormula (I)-(XXI) wherein X is a mono- or disaccharide, carboxylateddisaccharide, mono- or dicarboxylic acids, a salicylate, salicylic acid,aspirin, lactobionic acid, maltose, an amino acid, glycine, acetic acid,butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fattyacid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, atrisacharide, or a di- or trisaccharide monomer unit of heparin, heparansulfate, keratin, keratan sulfate, chondroitin, chondroitin-6-sulfate,chondroitin-4-sulfate, dermatin, dermatan sulfate, dextran, hyaluronicacid, glycosaminoglycan, or polypyranose.

Examples of suitable divalent groups forming the optional bridging groupY are straight- or branched-chain alkylene, e.g. of 2 or more,preferably 4 to 18 carbon atoms, —CO-alkylene-CO, —NH-alkylene-NH—,—CO-alkylene-NH—, cycloalkylene, wherein alkylene in each instance, isstraight or branched chain and contains 2 or more, preferably 2 to 18carbon atoms in the chain, —(—O—CH(CH₃)CH₂—)_(x)— wherein x is aninteger of 1 or more.

In another embodiment, in addition to the traditional phospholipidstructure, related derivatives for use in this invention arephospholipids modified at the C1 or C2 position to contain an ether oralkyl bond instead of an ester bond. These derivatives are exemplifiedhereinabove by the general formulae (VIII) and (IX).

In one embodiment of the invention, X is covalently conjugated to alipid. In another embodiment, X is covalently conjugated to a lipid viaan amide bond. In another embodiment, X is covalently conjugated to alipid via an esteric bond. In another embodiment, the lipid isphosphatidylethanolamine.

In one embodiment, cell surface GAGs play a key role in protecting cellsfrom diverse damaging agents and processes, such as reactive oxygenspecies and free radicals, endotoxins, cytokines, invasion promotingenzymes, and agents that induce and/or facilitate degradation ofextracellular matrix and basal membrane, cell invasiveness, white cellextravasation and infiltration, chemotaxis, and others. In addition,cell surface GAGs protect cells from bacterial, viral and parasiticinfection, and their stripping exposes the cell to interaction andsubsequent internalization of the microorganism. Enrichment of cellsurface GAGs would thus assist in protection of the cell from injuriousprocesses. Thus, in one embodiment of the invention, PLA2 inhibitors areconjugated to GAGs or GAG-mimicking molecules. In another embodiment,these compounds for use in the present invention provide wide-rangeprotection from diverse injurious processes, and are effective inamelioration of diseases that requires cell protection from injuriousbiochemical mediators.

In another embodiment, a GAG-mimicking molecule may be, inter alia, anegatively charged molecule. In another embodiment, a GAG-mimickingmolecule may be, inter alia, a salicylate derivative. In anotherembodiment, a GAG-mimicking molecule may be, inter alia, a dicarboxylicacid.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from an eye disease,including a lipid or phospholipid moiety bonded to a physiologicallyacceptable monomer, dimer, oligomer, or polymer; and a pharmaceuticallyacceptable carrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from an eye disease,including any one of the compounds for use in the present invention orany combination thereof; and a pharmaceutically acceptable carrier orexcipient. In another embodiment, the compounds for use in the presentinvention include, inter alia, the compounds represented by thestructures of the general formulae as described hereinbelow: (A), (I),(II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (IXa), (IXb), (X),(XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX),(XXI), (XXII), or any combination thereof.

The combination of lipids, such as, but not limited tophosphatidylethanolamine and phosphatidylserine, with additional monomeror polymer moieties, is thus a practical route to the production of newdrugs for medical purposes, provided that the resultant chemicalcomposition displays the desired range of pharmacological properties. Inone embodiment, the compounds for use in the present invention possess acombination of multiple and potent pharmacological effects in additionto the ability to inhibit the extracellular form of the enzymephospholipase A2. While the pharmacological activity of the compoundsfor use in the present invention described herein may be due in part tothe nature of the lipid moiety, the multiple and diverse combination ofpharmacological properties observed for the compounds for use in thepresent invention emerges from the ability of the compound structure toact essentially as several different drugs in one chemical entity.

In the cases described herein, the diversity of biological activitiesand the effectiveness in disease exhibited by the compounds for use inthe present invention far exceed the properties anticipated by use ofthe starting materials themselves, when administered alone or incombination. However, the phospholipid conjugate compounds, alone or incombination, are valuable when used in the methods of treating diseasesand conditions specifically described herein.

Eye Devices

It is to be understood that the compounds for use in the presentinvention may also be used in combination with any device which isapplied to an eye surface or applied to the internal regions of the eye.In one embodiment, such a device is a contact lens, while in otherembodiments, it is a corneal prosthetic device, prosthetic iris implant,scleral lens prosthetic device, an intra-ocular implant, a scleralbuckle, ophthalmic tantalum clip, ophthalmic conformer, artificial eye,absorbable implant, eye sphere implant, extraocular orbital implant,keratoprosthesis, intraocular lens, scleral shell, eye valve implant, ora combination thereof.

In one embodiment, the present invention provides a substrate having acoating on at least a portion of a surface of said substrate, saidcoating comprising a lipid or phospholipid moiety bound to aphysiologically acceptable monomer, dimer, oligomer, or polymer as anyof the embodiments describe hereinabove. In one embodiment, thephysiological acceptable monomer, dimer, oligomer, or polymer is apolypyranose. In one embodiment, the substrate is a contact lens. In oneembodiment, the substrate is an implant. In one embodiment, thesubstrate is part of a device for ophthalmic or ophthamologic use.

In another embodiment, this invention provides a contact lens solutioncomprising the compounds of the instant invention. Contact lenssolutions of the instant invention may comprise, inter alia, rewettingdrops, cleaning solutions, washing solutions, storage solutions, packingsolutions, saline solution, daily cleaner, multipurpose solution,hydrogen peroxide solution, or a combination thereof. Any solution whichmay be used for storage, preservation, or cleaning of a contact lens orlenses is considered to be an embodiment of this invention. The safetyand tolerability of solutions comprising the subject compounds ascomfort ingredients, for example, in contact lens packaging solutions,is exemplified in Example 6. In one embodiment, appliances for use withthe lenses and/or solutions of this invention may be coated with thecompounds for use in the instant invention as described herein, as well.

In one embodiment, contact lens solutions of the instant invention willadditionally comprise surfactants, tonicity agents, viscosity builders,anti-microbials, buffering agents, or a combination thereof. In oneembodiment, surfactants may be non-ionic, and in one embodiment maycomprise polyoxyethylene) and poly(oxypropylene), polyethylene glycolesters of fatty acids, e.g. coconut, polysorbate, polyoxyethylene orpolyoxypropylene ethers of higher alkanes (C12-C18). Examples includeTween® 20 (polysorbate 20) and Tween® 80, polyoxyethylene (23) laurylether (Brij® 35), polyoxyethyene (40) stearate (Myrj® 52),polyoxyethylene (25) propylene glycol stearate (Atlas® G 2612).

An amphoteric, cationic or anionic surfactant may also be present in thecontact lens solution. Amphoteric surfactants suitable for use in acomposition according to the present invention include materials of thetype are offered commercially under the trade name “Miranol”. Anotheruseful class of amphoteric surfactants are exemplified bycocoamidopropyl betaine commercially available under the trade nameAmphoso CA. Surfactants suitable for use in the invention can be readilyascertained, in view of the foregoing description, from McCutcheon'sDetergents and Emulsifiers, North American Edition, McCutcheon Division,MC Publishing Co., Glen Rock, N.J. 07452 and the CTFA InternationalCosmetic Ingredient Handbook, Published by The Cosmetic, Toiletry, andFragrance Association, Washington, D.C.

The pH of some contact lens solutions should, in one embodiment, bemaintained within the range of about 6.0 to 8.0, preferably about 6.5 to7.8. Suitable buffers may be added, such as boric acid, sodium borate,potassium citrate, citric acid, sodium bicarbonate, TRIS and variousmixed phosphate buffers (including combinations of Na₂HPO₄, NaH₂PO₄ andKH₂PO₄) and mixtures thereof. Generally, buffers will be used in amountsranging from about 0.05 to 2.5 percent by weight, and preferably, from0.1 to 1.5 percent. In one embodiment, the contact lens solutions ofthis invention contain a borate buffer, comprising one or more of boricacid, sodium borate, potassium tetraborate, potassium metaborate ormixtures of the same. Also, various buffer systems such as citrate,phosphate (appropriate mixtures of Na₂HPO4, NaH₂PO4, and KH₂PO4),bicarbonate, tromethamine and other appropriate nitrogen-containingbuffers (such as ACES, BES, BICINE, BIS-Tris, BIS-Tris Propane, HEPES,HEPPS, imidazole, MES, MOPS, PIPES, TAPS, TES, Tricine) can be used toensure a physiologic pH between about pH 6.5 and 8.5.

In one embodiment, the contact lens solutions of the present inventionare also adjusted with tonicity agents, to approximate the osmoticpressure of normal lacrimal fluids which is equivalent to a 0.9 percentsolution of sodium chloride or 2.5 percent of glycerol solution. Thesolutions are made substantially isotonic with physiological saline usedalone or in combination. In another embodiment, propylene glycol,lactulose, trehalose, sorbitol, mannitol or other osmotic agents mayalso be added to replace some or all of the sodium chloride.

Examples of suitable tonicity-adjusting agents include, but are notlimited to: sodium and potassium chloride, dextrose, glycerin, calciumand magnesium chloride. These agents are typically used individually inamounts ranging from about 0.01 to 2.5% (w/v) and preferably, from about0.2 to about 1.5% (w/v). Preferably, the tonicity agent will be employedin an amount to provide a final osmotic value of 200 to 400 mOsm/kg, andmore preferably between about 250 to about 350 mOsm/kg, and mostpreferably between about 280 to about 320 mOsm/kg.

It may also be desirable to optionally include water-soluble viscositybuilders in the solutions of the present invention. Because of theirdemulcent effect, viscosity builders have a tendency to further enhancea lens wearer's comfort by means of a film on the lens surfacecushioning impact against the eye. Included among the water-solubleviscosity builders are polymers like polyvinylalcohol cellulose-derivedpolymers, and povidone. In another embodiment, viscosity builderscomprise polyethylene glycol, surfactants, polyvinylpyrrolidone,polyvinyl alcohol, carboxymethyl cellulose and similar materials. Suchpolymers may be used in an amount of from about 0.01 to about 4.0 weightpercent or less. Surface-active agents, such as polysorbates,polyoxyethylenes and certain phosphonates, may be added to ensure properwetting and/or cleaning. Sequestering agents such asethylenedianiinetetraacetic acid (EDTA), phosphonates, citrate,gluconate and tartarate, are also common additives to preservatives,disinfection or cleaning solutions.

In another embodiment, nitrogen (non-fluorocarbon) is used as apropellant in the contact lens solution.

In another embodiment, the contact lens solution of the instantinvention further comprises anti-microbial compounds, which in oneembodiment, comprise glycosides, alkaloids, phenolics (anthocyanins,quinones, flavonols and flavonoids, etc.), terpenoids (includingphytosterols and carotenoids), or a combination thereof. In anotherembodiment, anti-microbial compounds comprise allicin, aucubin,berberine, bilberry extract, caffeic acid, chlorogenic acid, Echinaceaextract, ferulic acid, hydrastine, lipoic acid, naringin, oleuropein,proanthocyanidins, quercetin, rutin, or a combination thereof, which inone embodiment are present in amounts of 10 to 10,000 parts per million.In another embodiment, saponins, can be used as natural plantsurface-active or cleaning agents in lens solutions. Specifically,triterpenoid saponins and steroid saponins are particularly effective incontact lens or ophthalmic solutions. In another embodiment, thesolution comprises benzyldimethyl{2-[2-(p-1,1,3,3-tetramethylbutylphenoxy)ethoxy]ethyl}ammonium chloride(BDT) as an anti-microbial compound.

In another embodiment, a contact lens solution of the instant inventioncomprises preservatives, which in one embodiment comprise thimerosal,edetate disodium, sorbic acid, polyaminopropyl biguanide, POLYQUAD(polyquartenium-1), EDTA, or a combination thereof.

In one embodiment, a contact lens solution may be a daily cleaner, whichin one embodiment comprises cocoamphocarboxyglycinate, sodium laurylsulfate, hexylene glycol, sodium chloride, sodium, Tween 21, microlenspoloxamer 407, potassium chloride, poloxamine, isopropyl alcohol,amphoteric 10, or a combination thereof.

In one embodiment, contact lens solution may comprise an enzyme, whichin one embodiment may be pancreatin, papain, subtilisin, or acombination thereof, which in one embodiment may be for removing proteinfrom the lens.

In another embodiment, a device for use with contact lenses may comprisecompounds for use in the instant invention. In one embodiment, such adevice may be a standard device commercially available for the storage,cleaning, disinfection, and/or carriage of contact lenses, which in oneembodiment, is characterized by a hollow well. In one embodiment, asingle device may be suitable for storage, cleaning, disinfection,and/or carriage of contact lenses, while in another embodiment, each isa separate device.

In one embodiment, the substrate, which in one embodiment is a contactlens, contact lens device, contact lens solution and/or intraoculardevice, comprising compounds for use in the instant inventionsuppresses, inhibits, prevents or treats eye-related disorders,including, inter alia, those described hereinabove, in a subject. Inanother embodiment, the substrate comprising compounds for use in theinstant invention prevents or treats proteinaceous deposits accumulatingon the substrate. In another embodiment, the compounds for use in theinstant invention endow the surface of the substrate with the propertyof being more hydrophilic, which in one embodiment, may increasecomfort, decrease eye dryness, or a combination thereof. In anotherembodiment, the compounds for use in the instant invention preventadverse reactions that are directly or indirectly related to thesubstrate, such as corneal edema, inflammation, or lymphocyteinfiltration. In another embodiment, the substrate comprising compoundsfor use in the instant invention increases wettability, decreasesadhesion, increases biocompatability, provides UV shielding, preventsglare, decreases dryness, grittiness, general discomfort, preventsmicrobial (in one embodiment, bacterial) infections, or a combinationthereof or provides other desirable characteristics and properties tothe substrate that are known in the Art.

Preparation of Compounds for Use in the Present Invention

In one embodiment, the preparation of high molecular weight compoundsfor use in the methods of the present invention is as described in U.S.Pat. No. 5,064,817, which is incorporated fully herein by reference. Inone embodiment, these synthetic methods are applicable to thepreparation of low molecular weight compounds for use in the presentinvention as well, i.e. compounds for use in the present inventioncomprising monomers and dimers as the conjugated moiety, withappropriate modifications in the procedure as would be readily evidentto one skilled in the art. The preparation of some low molecular weightcompounds for use in the present invention may be conducted usingmethods well known in the art or as described in U.S. patent applicationSer. No. 10/952,496, which is incorporated herein by reference in itsentirety.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present invention toits fullest extent. The following preferred specific embodiments are,therefore, to be construed as merely illustrative, and not limiting theremainder of the disclosure in any way whatsoever.

EXAMPLES

The abbreviations used in the examples below are:

PE=phosphatidyl-ethanolamine

HA=hyaluronic acid

Cpd=Compound

Cpd XXII=dipalmitoyl-PE conjugated to HA

Cpd XXIII=dimyristoyl-phosphatidyl-ethanolamine linked to HA

Cpd XXIV=PE conjugated to heparin

Cpd XXV=PE conjugated to chondroitin sulfate A (CSA)

Cpd XXVI=PE conjugated to carboxymethyl cellulose (CMC)

Cpd XXVII=PE conjugated to Polygeline (haemaccel)

Cpd XXIX=PE conjugated to dextran

Cpd XXX=PE conjugated to aspirin

Cpd LXXXVIII=PE conjugated to glutaryl

Cpd LI=PE conjugated to alginate

The compounds used in the examples below were prepared as described inU.S. patent application Ser. No. 10/952,496, which is fully incorporatedherein by reference.

Example 1 Effect of Lipid Conjugates in an In Vitro Model of DiabeticRetinopathy Human Retinal Endothelial Cell Culture

Human eyes from donors are obtained and human retinal endothelial cells(HRECs) are isolated. The identity of HRECs is validated bydemonstrating endothelial cell incorporation of fluorescence-labeled,acetylated LDL, and by fluorescence-activated cell-sorting analysis. Todetermine the effect of high glucose, HRECs are grown for 7 days innormal (5.5 mM) or high (25 mM) D-glucose medium.

Bovine Retinal Endothelial Cell Culture

Isolated bovine retinas are homogenized in ice-cold Eagle's minimalessential medium (MEM) with HEPES by a Teflon-glass homogenizer andmicrovessels trapped on an 83 mm nylon mesh. Vessels are transferredinto 2× MEM containing 500 μg/ml collagenase, 200 μg/ml pronase (BDH,Poole, UK) and 200 μg/ml DNase at 37° C. for 20 min The resultant vesselfragments are trapped on 53 μm mesh, washed with cold MEM, andcentrifuged at 225×g for 10 min. The pellet is suspended inmicrovascular endothelial cell basal medium (MECBM) with growthsupplement (TCS Works Ltd., Buckingham, UK) at 37° C., 5% CO₂ for 3days. Confluent cells are used between passages 1 and 3.

ELISA for VEGF

VEGF protein concentration is determined from retinal endothelial cellculture-conditioned medium using the Quantikine® Human VEGF ImmunoassayELISA kit (R & D Systems). Retinal endothelial cell culture are treatedwith either 5.5 or 25 mM glucose and aliquots are taken daily foranalysis.

ELISA for IGF-I

IGF-I protein concentration is determined from retinal endothelial cellculture-conditioned medium using the Quantikine® Human IGF ImmunoassayELISA kit (R & D Systems). Retinal endothelial cell culture are treatedwith either 5.5 or 25 mM glucose and aliquots are taken daily foranalysis.

Statistical Analysis

Data are analyzed using the Student's t-test and reported asmean±standard deviation (SD). A p value <0.05 is considered significant.

Treatment with 1, 5, and 10 μM, and other concentrations of CompoundsXXII, XXIII, or XXV restore the levels of IGF-1 and VEGF in human andbovine retinal endothelial cell culture to the level of controls.

In another embodiment, other markers may be examined including ICAM-1,VCAM-1, HIF-1, transmembrane reductase (TMR), and EPO; pigmentepithelium-derived factor (PEDF) in the eye, markers of oxidative stressincluding osmotic stress after accumulation of sorbitol, increasedcytosolic NADH/NAD ratio, depletion of NADPH and accumulation offructose with the resulting non-enzymatic production of advancedglycation end products (AGES); and/or ESR, fibrinogen, SDF/1_, RANTES,EpOx, Haptoglobin and ACE in peripheral blood. Antibodies are used toprobe the GAG portion of the conjugate over a time course compared tounconjugated control, and show greater local persistence. In oneembodiment, GAGs are tagged.

Example 2 Effect of Lipid Conjugates in an In Vivo Model of DiabeticRetinopathy

Diabetes is induced Long-Evans rats via ip streptozotocin (STZ)injections at 70-85 mg/kg, for 3 to 5 days. To help ease the transitionto diabetes, the rats are given 10% sugar water for 24 hours post-STZinjection. Retinal photographs are taken and blood glucose tests areperformed to determine baselines for each rat. The normal glucose rangefor a rat is 80-100 mg/dl.

Alternatively, C57BL/6 mice (SLC, Shizuoka, Japan) are used. Postnatalday (P)7 mice with their nursing mothers are maintained for a full 5days in 80% oxygen to generate the nonvascular retinal area. On P12,they are placed in normoxia for an additional 5 days to induce retinalneovascularization.

Each animal is glucose-tested and photographed with a fundus camera on aweekly basis to record the progression of diabetic retinopathy.Approximately 20-30 minutes prior to starting pictures, one drop of 1%atropine is placed in each eye. The rats are anesthetized with sodiumpentobarbital, at a dose of 60 mg/kg, to keep them immobile, and theninjected ip with 0.1 ml of 25% fluorescein, which is used to visualizethe retinal blood vessels when illuminated by blue light, and todetermine the relative leakage of blood by the intensity in thebackground. The retinal leakage score is determined by digital analysisof the vascular and extravascular fluorescence.

Rats treated with PTZ show significantly increased retinal leakagescores compared to vehicle-treated controls. Treatment with 1, 5, and 10μM or other concentrations of Compounds XXII, XXIII, or XXV decrease theretinal leakage score of PTZ-treated rats back to the level of controls.

After 14 days, rats are sacrificed and their retinas examined forretinal mRNA and protein levels of intercellular adhesion molecule(ICAM)-1, vascular endothelial growth factor (VEGF) by RT-PCR and ELISA.

RT-PCR for Intercellular Adhesion Molecule-1 and Vascular EndothelialGrowth Factor Receptor-1 and -2

Total RNA is isolated from the retina using extraction reagent (Isogen;Nippon Gene, Toyama, Japan) and reverse-transcribed with a cDNAsynthesis kit (First-Strand; Pharmacia Biotech, Uppsala, Sweden)according to the manufacturer's protocols. PCR is performed with Taq DNApolymerase (Toyobo, Tokyo, Japan) in a thermal controller (MiniCycler;MJ Research, Watertown, Mass.). The primer sequences are as follows:5′-ATG TGG CAC CAC ACC TTC TAC AAT GAG CTG CG-3′ (sense) and 5′-CGT CATACT CCT GCT TGC TGA TCC ACA TCT GC-3′ (antisense; 37 bp) for B-actin,5′-GTG TCG AGC TTT GGG ATG GTA-3′ (sense) and 5′-CTG GGC TTG GAG ACT CAGTG-3′ (antisense; 505 bp) for mouse intercellular adhesion molecule(ICAM)-1. Human/mouse vascular endothelial growth factor receptor-1(VEGF R1) primers (302 bp; PCR Primer Pair; R&D Systems, Inc.,Minneapolis, Minn.) and human/mouse VEGF R2 primers (569 bp; PCR PrimerPair; R&D Systems, Inc.) are used for VEGFR-1 and -2, respectively.

ELISA for ICAM-1 and VEGFR-1 and -2

The animals are killed with an overdose of anesthesia, and the eyes areimmediately enucleated. The retina is carefully isolated and placed into200 μL lysis buffer (0.02 M HEPES, 10% glycerol, 10 mM Na₄P₂O₇, 100 μMNa₃VO₄, 1% Triton, 100 mM NaF, 4 mM EDTA [pH 8.0]) supplemented withprotease inhibitors, and sonicated. The lysate is centrifuged at 15,000rpm for 15 minutes at 4° C., and the ICAM-1 and VEGFR-1 and -2 levels inthe supernatant are determined with mouse ICAM-1 and VEGFR-1 and -2 kits(Techne Corp., Minneapolis, Minn.) according to the manufacturer'sprotocol. The tissue sample concentration is calculated from a standardcurve and corrected for protein concentration.

Treatment with 1, 5, and 10 μM and other concentrations of CompoundsXXII, XXIII, or XXV dose-dependently decreases the levels of ICAM-1 andVEGF in PTZ-treated rats back to the level of controls.

Example 3 Effect of Lipid Conjugates in Patients with DiabeticRetinopathy

Plasma samples are collected from Type I diabetic patients. Their gradeof retinopathy is characterised according to a modified Airlie housetechnique: (a) no retinopathy (n=6), (b) background retinopathy (n=10),(c) proliferative retinopathy (n=6), and (d) advanced proliferativeretinopathy requiring vitrectomy (n=16). Plasma samples fromnon-diabetic age-matched control subjects are also collected. Vitreoussamples are collected from patients with advanced proliferative diabeticretinopathy prior to undergoing vitrectomy. Research Ethics Committeeapproval and informed consent are obtained from all patients.

Plasma and Vitreous Samples

Venous blood samples are collected from the patients and controlsubjects. Plasma is harvested by centrifugation, aliquoted and stored at−70° C. Approximately 0.5-1 ml of undiluted vitreous fluid is collectedfrom the eye prior to irrigation of the vitreous, transported on dry iceand stored at −70° C.

Enzyme-Linked Immunosorbent Assay (ELISA) for CD105

White 96-well micro-titre plates are coated with anti-CD105 Mab E9 (100μl/well) diluted 1/1000 in 0.1 M PBS, and incubated in a humidifiedchamber overnight at 4° C. The coated plates are blocked using 1% BSAand 0.1% Tween 20 in 0.1 M PBS (PBS-Tween) for 2 h at room temperature.Test samples, 1/2 diluted in PBS-Tween, are added to the plates induplicate. Plasma with pre-determined CD105 (100 ng/ml) is titrated tomake a standard curve in each plate. After overnight incubation at 4°C., biotinylated Mab E9 (1/2000 dilution), 100 μl/well, is added to theplates, followed by incubation at 4° C. in a humidified chamber for 3 h.HRP-conjugated avidin at 1/2000 dilution in PBS-Tween and 1% BSA isadded(100 μl/well), and plates are incubated at room temperature for 30min Three washes with PBS-Tween are carried out between each of theprocedures. Finally, 100 μl/well of Amerlite signal reagent (AmershamUK) are added to each well and light emission is measured immediately at420 nm in an Amerlite plate reader (Kodak Clinical Diagnostics,Aylesbury, UK).

Indirect Immunoassay for VEGF

White 96-well plates are coated with 1000/well of goat anti-VEGF-165antibody (R&D systems), diluted 1/1000 (1 μg/ml) in 0.1M carbonatebuffer (pH 9.6), and incubated in a humid box overnight at 4° C. Thecoated plate is blocked with 1% (w/v) bovine serum albumin (BSA), 0.01%(v/v) Tween 20 in 0.1 M PBS (PBSTween) for 2 h at room temperature.Serum samples are added in duplicate to the plates (100 μl/well, diluted1/2 in PBS-Tween). A standard curve is generated using recombinant humanVEGF (R & D systems) in a range of 0.1-40 ng/ml on each plate. Afterovernight incubation at 4° C., rabbit anti-VEGF antibody (Santa CruzBiotechnology) is added (100 μl/well) to the plate at 1/2000 dilution (1μg/ml) in PBS-Tween and incubation is carried out for 3 h at 4° C. Thisis followed by the addition of HRP-conjugated goat anti-rabbit antibody(0.5μg/ml) (diluted 1/2000 with 1%BSA in PBSTween), and additionalincubation with shaking for 30 min at room temperature. Three washeswith PBS-Tween are carried out between each of the steps. Finally, 100μl/well of Amerlite chemiluminescence signal reagent are added and theplate is read immediately in a plate reader. The measured values oflight emission are converted into absolute concentration by reference tothe VEGF standard curve.

Treatment with 1, 5, and 10 μM and other concentrations of CompoundsXXII, XXIII, or XXV dose-dependently decreases the plasma levels ofCD105 and vitreous levels of VEGF in patients with diabetic retinopathy.In addition, fluorescein angiography, retinal photography, andultrasound imaging of the eye are used to evaluate progression of thedisease.

Example 4 Effect of Contact Lens Solution Comprising Lipid Conjugates onProtein and Lipid Deposition

Contact lenses are exposed to both a protein and lipid artificialdeposition solution (ATS) in order to assess both the deposit inhibitionof the contact lens solution of the instant invention compared to asolution known in the Art, such as ReNu® Rewetting Drops which contains0.10% poloxamine, 0.50% boric acid, 0.35% sodium borate, 0.40% sodiumchloride, 0.10% EDTA, and 0.15% sorbic acid. To test for depositinhibition, lenses are preconditioned with the solution of the instantinvention by soaking the lens in the solution for one hour prior todeposition. After deposition and incubation, the lenses are rinsed with0.9% saline solution (without sorbic acid).

A. Protocol for Testing Protein Deposit Inhibition:

For preparation of the standards, unworn contact lenses are taken out oftheir vials, left to air-dry and then placed in glass test tubes alongwith standard BSA solution. An in vitro protein mixture consisting oflysozyme, lactoferrin, human serum albumin and mucin in MOPS buffer isused. The pH of the solution is adjusted to 7.2 using 1 N HCl and anosmolality equal to 326 mOsm. After one hour of pre-soaking, the lensesare removed from the formulation and placed in 1.5 ml of the proteinmix. The lenses are then incubated in the protein mix at 37° C. in ashaking water bath for 48 hours. Protein analysis is done using thecolorimetric BCA analytical method (Sigma). The method employs theprotein induced reduction of Cu(II) to Cu(I). A purple complex (Amax=562nm) is formed following the addition of bicinchoninic acid (BCA) to thereduced copper. The intensity of the complex is directly proportionalover the protein concentration range of 5 μg/ml to 2000 μg/ml. Followingincubation at 37° C., the rate of color development is slowedsufficiently to allow large numbers of samples to be assayed in a singlerun. The standard protein solution utilized is BSA with a standardconcentration range of 0 to 50 μg. Two mls of a mixture of bicinchoninicacid (BCA) and Cu(II) sulfate is added to each test tube, which are thenvortexed. Tubes are then covered and placed in a water bath at 37° C.for 15 minutes. After incubation, the purple complex develops. Samplesand standards are read in a spectrophotometer at 562 nm Proteinconcentration is determined from a standard plot of absorbency vs.concentration (ig).

B. Protocol for Testing Lipid Deposit Inhibition:

Seven contact lenses per test solution are preconditioned with therespective test formulations by soaking the lenses in the formulationfor one hour. The lenses are removed from the formulation, and placed in1.5 mls of a lipid mix (palmitic acid methyl ester (PAME), cholesterol,squalene and mucin in MOPS buffer). Mucin is utilized as a surfactant toaid in the solubilization of the lipids. Lenses are then incubated inthe lipid mix at 37° C. in a shaking water bath for 24 hours. Afterincubation, the lenses are removed from the test solution and rinsedwith physiological saline solution (without sorbic acid) to remove anyresidual deposition solution. Lenses are then placed in glass vials forextraction. A three-hour 1:1 CHCl₃/MeOH extraction is subsequentlyfollowed by a three-hour hexane extraction. Extracts are then combinedand run on a Hewlett Packard GC. The column utilized is an HP-Ultra 1with an FID detector and He as the carrier gas. Standard solutions ofeach of the lipids in the deposition mix are made in 1:1 CHCl₃/MeOH andthe concentration of lipid extracted from the lenses is determined

C. Results:

The protein and lipid deposition values for the contact lensespre-soaked in control solution provide a baseline with which to assessthe potential cleaning efficacy and deposit inhibition attributes ofeach of the formulations tested. The contact lens solution of theinstant invention inhibits both lipid and protein deposition, indicatingthat the test formulations are coating the lens in such a way as tohinder lipid and protein uptake.

Example 5 Safety and Tolerability of Contact Lens Solution ComprisingLipid Conjugates

Twenty (20) subjects are enrolled in a 4-hour non-dispensing studycomparing the contact lens solution comprising lipid-conjugates tocontact lens solution comprising unconjugated GAGs or to ReNu® RewettingDrops. The subjects are all habitual soft spherical contact lenswearers. Their mean spherical Rx's are determined for test and controleyes. Each subject wears a pair of contact lenses for approximately 4hours. The eye receiving the test solution is randomly selected andremains constant for the duration of the study. Subjects are asked toplace two drops of each solution into the appropriate eye every houruntil the four-hour visit. The subjects and investigator are blinded tosolution identity. Prior to lens insertion, a spherical refraction isperformed through which high contrast visual acuity with high ambientillumination (HCHI) is measured. Corneal and conjunctival staining andlimbal and bulbar injection are assessed with the slitlamp. Each subjectis then fitted with a pair of contact lenses of their prescription. Eachlens is evaluated for centration and movement, comfort, anddeposits/wettability. A spherical over-refraction is then performed. Theendpoint of the over-refraction is compared to the refractive endpointto determine the apparent “on-eye” lens power. LogMAR visual acuityunder HCHI testing conditions is measured through the over-refraction.Finally, two drops of each solution are instilled into the appropriateeyes, and the subject is asked to rate any sting/burn and the amount.Testing is repeated at the four-hour visit in reverse order, exceptwithout repeating the baseline refraction. A two-way ANOVA incorporatingTime and Solution is used to test for differences in each of theparametric dependent variables measured. Non-parametric data areanalyzed by Friedman ANOVA. Differences at the p<=0.05 level areconsidered to be statistically significant.

Subjects are evaluated for the effects of the solution comprisinglipid-conjugates for comfort, apparent lens Rx power, and sting/burnvisual analog score (i.e. lower sting/burn), lens movement/centration,and anterior ocular physiology in the eye treated with lipid-conjugatesolutions compared to the eye treated with control solution.

Example 6 Safety and Tolerability of Contact Lens Packaging SolutionComprising Lipid Conjugates Materials and Methods

Contact lenses were exposed to a solution comprising lipid conjugates inorder to assess their efficacy as a comfort ingredient in a contact lenssolution. Five lipid conjugate compounds, Cpd LI (AlgPE:120), Cpd XXV(CSAPE:120), Cpd XL (HemPE:75), Cpd XXVIII (HesDMPE:90 (HetaStarch)) andCpd XXVI (CMPE:75), were tested.

Cpd LI, Cpd XXV, Cpd XL, Cpd XXVIII and Cpd XXVI were tested for theirsolubility in an aqueous solution containing Na₂HPO₄×7H₂O, NaH₂PO₄ andNaCl, pH 7.3, with an osmolality of 250 mOsm/kg.

PV lenses were tested for lens compatibility and stability. Each lenswas soaked in a glass vial containing 3 ml of a solution of one of thefollowing: Cpd XXV, Cpd XL, Cpd XXVIII or Cpd XXVI. The glass vials wereautoclaved at 121-123° C. for 30 min The lenses and the solutions wereexamined for optical parameters and physical appearance immediatelyfollowing autoclaving and again after one month at 40° C.

Cytotoxic effects of the compounds XXV, XL, XXVIII and XXVI on kidney(MDCK) and SV40 human corneal (HCEC) epithelial cells were evaluated.Each of the compounds in solution was added to MDCK and HCEC cellculture models, with physiological saline or HBSS serving as a control.The compounds were individually tested for their toxicity to L929 mousefibroblasts in an agar diffusion model, in which cells were separatedfrom each compound by a layer of agar. In this test only compounds thatcan diffuse through the agar and are toxic to the cells are detected.The L929 monolayer cultures were incubated with the compounds, andobserved for cytotoxicity.

PV lenses were soaked in solutions of the compounds and tested. In vivoevaluation was assessed in a one-day rabbit ocular irritationexperiment. PV lenses were soaked in solutions of the compounds andplaced on the corneas of test rabbits.

Results

Cpds XXV, XL and XXVIII were soluble. Cpd XXVI was soluble uponautoclaving the solution. Cpd LI was insoluble.

The optical parameters and physical appearance of the contact lensesimmediately following autoclaving were within industry specifications.There were no significant changes in the pH or osmolality of any of thesolutions. A follow-up check one month later also revealed nosignificant changes in either lens or solution parameters.

Results with the compounds on MDCK and HCEC paralleled those of thecontrols. No cytotoxicity to L929 monolayer cultures was noted.

No or low levels of ocular irritation were observed with the PV lensesin vivo. There was no statistical difference between treated vs. controlPV lenses.

It will be appreciated by persons skilled in the art that the presentinvention is not limited by what has been particularly shown anddescribed herein above and that numerous modifications, all of whichfall within the scope of the present invention, exist. Rather, the scopeof the invention is defined by the claims which follow:

What we claim is:
 1. A method of treating a disease or disorder of theeye in a subject comprising the step of contacting said subject with acompound comprising a lipid or phospholipid moiety bound optionally viaa spacer to a physiologically acceptable polymer, and/or apharmaceutically acceptable salt or a pharmaceutical product thereof. 2.The method according to claim 1, wherein said phospholipid moiety isphosphatidylethanolamine.
 3. The method according to claim 2, whereinsaid phosphatidylethanolamine is dipalmitoyl phosphatidylethanolamine.4. The method according to claim 2, wherein saidphosphatidylethanolamine is dimyristoyl phosphatidylethanolamine
 5. Themethod according to claim 1, wherein said physiologically acceptablemonomer, dimer, oligomer, or polymer is polygeline.
 6. The methodaccording to claim 1, wherein said physiologically acceptable monomer,dimer, oligomer, or polymer is a polypyranose.
 7. The method accordingto claim 6, wherein said polypyranose is carboxymethylcellulose.
 8. Themethod according to claim 6, wherein said polypyranose is alginate. 9.The method according to claim 6, wherein said polypyranose ishydroxyethyl starch.
 10. The method according to claim 1, wherein thelipid or phospholipid moiety bound optionally via a spacer to aphysiologically acceptable monomer, dimer, oligomer, or polymer via anester or amide bond, and/or a pharmaceutically acceptable salt or apharmaceutical product thereof is represented by the structure of thegeneral formula (A):

wherein L is a lipid or a phospholipid; Z is either nothing,ethanolamine, serine, inositol, choline, phosphate, or glycerol; Y iseither nothing or a spacer group ranging in length from 2 to 30 atoms; Xis a physiologically acceptable polymer; and n is a number from 2 to1000.
 11. The method of claim 10, wherein L is phosphatidyl, Z isethanolamine, Y is nothing, and X is carboxymethylcellulose or aglycosaminoglycan.
 12. The method of claim 10, wherein thephosphatidylethanolamine moiety is dipalmitoyl or dimyristoylphosphatidylethanolamine.
 13. The method of claim 10, wherein the lipidor phospholipid moiety bound optionally via a spacer to aphysiologically acceptable monomer, dimer, oligomer, or polymer via anester or amide bond, and/or a pharmaceutically acceptable salt or apharmaceutical product thereof is represented by the structure of thegeneral formula (I):

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; and Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; X is aphysiologically acceptable polymer; and n is a number from 1 to 1000.14. The method of claim 13, wherein n is a number from 2 to
 100. 15. Themethod of claim 10, wherein the lipid or phospholipid moiety boundoptionally via a spacer to a physiologically acceptable monomer, dimer,oligomer, or polymer via an ester or amide bond, and/or apharmaceutically acceptable salt or a pharmaceutical product thereof isrepresented by the structure of the general formula (III):

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Z is eithernothing, inositol, choline, or glycerol; Y is either nothing or a spacergroup ranging in length from 2 to 30 atoms; X is a glycosaminoglycan;and n is a number from 1 to
 1000. 16. The method of claim 15, wherein nis a number from 2 to
 100. 17. The method of claim 10, wherein the lipidor phospholipid moiety bound optionally via a spacer to aphysiologically acceptable monomer, dimer, oligomer, or polymer via anester or amide bond, and/or a pharmaceutically acceptable salt or apharmaceutical product thereof is represented by the structure of thegeneral formula (IV):

wherein R₁ is either hydrogen or a linear, saturated, mono-unsaturated,or poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Z is eithernothing, inositol, choline, or glycerol; Y is either nothing or a spacergroup ranging in length from 2 to 30 atoms; X is a glycosaminoglycan;and n is a number from 1 to
 1000. 18. The method of claim 17, wherein nis a number from 2 to
 100. 19. The method of claim 10, wherein the lipidor phospholipid moiety bound optionally via a spacer to aphysiologically acceptable monomer, dimer, oligomer, or polymer via anester or amide bond, and/or a pharmaceutically acceptable salt or apharmaceutical product thereof is represented by the structure of thegeneral formula (V):

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is either hydrogen or a linear, saturated, mono-unsaturated,or poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; Z is either nothing, inositol, choline, or glycerol; Y is eithernothing or a spacer group ranging in length from 2 to 30 atoms; X is aglycosaminoglycan; and n is a number from 1 to
 1000. 20. The method ofclaim 19, wherein n is a number from 2 to 100.